The aim of the present study was to develop enzyme linked immunosorbent assays (ELISAs) for detection of specific antibodies against <i>Babesia bovis</i> and <i>B.bigemina</i> for use in epidemiological studies of bovine babesiosis in Brazil. These tests were developed using purified proteins of each parasite, which were identified as species-specific in a comprehensive immunochemical characterisation of diffrent stocks of <i>Babesia</i> parasites. The review of the literature on bovine babesiosis covers the geographical distribution, transmission and life cycle, and <i>in vitro</i> cultivation of both species, as well as immunology, diagnosis, epidemiology and control of the disease and its current situation in Brazil. The thesis first describes a series of studies on the <i>in vitro</i> culture of <i>B.bovis</i>, which include the evaluation of the effect of different sera on <i>in vitro</i> growth, the incorporation of feeder cells (bovine aortic endothelial and mouse peritoneal cells) into cultures, cloning of one isolate by <i>in vitro</i> limiting dilutions, initiation of cultures from low parasitaemia blood, and several attempts to establish African isolates of <i>B.bigemina in vitro</i>. Methods for concentration of <i>Babesia</i> infected red blood cells and free parasites were then examined. These included the use of differential hypotonic lysis, density gradient centrifugation, induction of free merozoites into culture supernatant and techniques for fractionation of exoantigens present in culture supernatant, namely high performance liquid chromatography. Somatic components and exoantigens of different stocks of parasites were characterised immunochemically by Western immuno-blotting, and immunoprecipitation of <SUP>35</SUP>S-methionine labelled proteins by acrylamide gel electrophoresis, using a wide variety of sera which included calf sera experimentally raised against different stocks of each parasite, sera collected in the field in Brazil and a panel of monoclonal antibodies previously produced against <i>B.bovis</i> and characterised by IFAT, Western immuno-blotting and ELISA in the present study.
| Identifer | oai:union.ndltd.org:bl.uk/oai:ethos.bl.uk:660382 |
| Date | January 1993 |
| Creators | Passos, Lygia Maria Friche |
| Publisher | University of Edinburgh |
| Source Sets | Ethos UK |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Source | http://hdl.handle.net/1842/29934 |
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