Name :     Doaa Mohamed Abd El-Gayed

Date of Birth :     15/6/1979

Nationality :         Egyptian

Previous Degree : M.Sc.

Registration Date : 2004

Awarding Date : 11/2/2008

Supervisors :        Prof. Dr. Somaya Osman  El-Deeb

                             Dr. Azza Mohamed El-Amir

                             Dr. Maysa Ahmed Shalaby

Examiners :         Prof. Dr. Somaya Osman El-Deeb

Prof. Dr. Frank Wunderlich

          Prof. Dr. Mostafa Mahmoud Ramadan

Title of Thesis :

Immunolocalizaton of Antigenic Epitopes Expressed During Fasciola gigantica infection Using Polychlonal Antibodies in Cattles

Key Words : Immunolocalization – Fasciola gigantica – Polyclonal antibodies

Summary :

Although serological tests are available for the diagnosis of the fascioliasis, they are of generally low specificity because of cross- reactivity with antigens from other parasites. Several antigens are needed for efficient diagnosis of Fasciola. The most common one is the E/S antigens. The E/S products may not only have an important role in the induction of protective immunity but contain antigens useful for immuno-diagnosis of fascioliasis.

This study deals with "Immunlocaliztion of antigenic epitopes expressed during F. gigantica infection using polyclonal antibodies in cattle". In addition, it aims to screen all possible organs for the localization of E/S in an attempt to find out a suitable method for diagnosing the disease as early possible.

       To achieve these goals, E/S Fasciola antigens were preparede for raising of pAb in rabbits followed by Purification of anti- F. gigantica E/S IgG PAb. Localization of antigenic epitopes in different life cycle stages of F. gigantica, in PBMNCs and different organs of naturally infected cattles using anti- F. gigantica E/S IgG PAb was cdeveloped.

       Application of the immunoperoxidase techniques using anti- F. gigantica E/S IgG PAb prove the presence of antigenic epitopes in the different stages of  F. gigantica except eggs. Positive peroxidase reactions were mainly detected in the tegument and gut of miracidiae, cercariae, metacercariae and tail of cercariae. In addition, at the tegument, spine surface, muscularis and gut region of the adult worms were also positive.

Immunoperoxidase localization of E/S products by anti-F. gigantica E/S IgG pAb demonstrated that the E/S antigens were located in many organs of naturally infected cattle. Accompanying histopathological changes were studied microscopically at the same intervals using haematoxylin and eosin stained sections.

As regards the liver, The E/S Fasciola antigens were detected in the cytoplasm of the infected liver cells, histiocytes. Hepatic pathological changes appeared as loss of hepatic lobular architecture, damage to the hepatocytes. Heavy infiltration of eosinophils, lymphocytes and plasma cells in between hepatocytes and in portal area of the liver.

As regards thenfected gallbladder, The E/S Fasciola antigens were detected in muscular layer. Pathological changes appeared in the form of hyperplasia of the mucosal lining with focal erosion. In addition to the proliferation and hyperplasia of the lining epithelial cells of the bile duct, there is superficial focal erosion of the lining epithelium of small bile ducts in the portal tracts.

 

 

 

 

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