The aim of this thesis was to develop diagnostic and quantitative assays/tools based on western blotting, confocal microscopy and immunogold electron microscopy, as well as flow cytometry, to determine the localisation and potential function of the highly expressed cathepsin L of the myxozoan parasite Sphaerospora molnari (SmolCL) in its fish host, the common carp. Furthermore, a recombinant SmolCL was used in a vaccine trial to estimate its potential for raising antibodies in carp and test their immunoprotective potential towards S. molnari. This thesis provides new methodological tools for research and allows a greater understanding of myxozoan parasite-fish host interactions based on proteolytic enzymes.
Anotace v angličtině
The aim of this thesis was to develop diagnostic and quantitative assays/tools based on western blotting, confocal microscopy and immunogold electron microscopy, as well as flow cytometry, to determine the localisation and potential function of the highly expressed cathepsin L of the myxozoan parasite Sphaerospora molnari (SmolCL) in its fish host, the common carp. Furthermore, a recombinant SmolCL was used in a vaccine trial to estimate its potential for raising antibodies in carp and test their immunoprotective potential towards S. molnari. This thesis provides new methodological tools for research and allows a greater understanding of myxozoan parasite-fish host interactions based on proteolytic enzymes.
The aim of this thesis was to develop diagnostic and quantitative assays/tools based on western blotting, confocal microscopy and immunogold electron microscopy, as well as flow cytometry, to determine the localisation and potential function of the highly expressed cathepsin L of the myxozoan parasite Sphaerospora molnari (SmolCL) in its fish host, the common carp. Furthermore, a recombinant SmolCL was used in a vaccine trial to estimate its potential for raising antibodies in carp and test their immunoprotective potential towards S. molnari. This thesis provides new methodological tools for research and allows a greater understanding of myxozoan parasite-fish host interactions based on proteolytic enzymes.
Anotace v angličtině
The aim of this thesis was to develop diagnostic and quantitative assays/tools based on western blotting, confocal microscopy and immunogold electron microscopy, as well as flow cytometry, to determine the localisation and potential function of the highly expressed cathepsin L of the myxozoan parasite Sphaerospora molnari (SmolCL) in its fish host, the common carp. Furthermore, a recombinant SmolCL was used in a vaccine trial to estimate its potential for raising antibodies in carp and test their immunoprotective potential towards S. molnari. This thesis provides new methodological tools for research and allows a greater understanding of myxozoan parasite-fish host interactions based on proteolytic enzymes.
Prof. Vácha welcomed the student, committee members and the audience. The supervisor Dr. Holzer was connected online, Dr. Štěrbová was not present.
The student presented the student myxozoan parasite and infection cycle in the carp followed by the description of the studied protein. An experimental setup with a detailed description of the various methods and plenty of results were presented. In the end, the results were summarized and discussed. The supervisor and the opponent presented their reviews. The student mostly answered the questions. Then he answered questions from the committee on the application of vaccines.