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Title: Genotoxins in the marine environment
Author: Harvey, J. S.
Awarding Body: University College of Swansea
Current Institution: Swansea University
Date of Award: 1995
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This thesis describes the development and optimisation of the 32P-postlabelling assay and the Restriction Site Mutation (RSM) assay for the detection of DNA adducts and mutations, respectively, in the marine bioindicator species, Mytilus spp. The standard approach to adduct analysis was modified, to eliminate various artefacts connected directly to the 32P-postlabelling procedure and to the method of DNA isolation. Initial studies demonstrated that specimens of Mytilus spp. from several locations possessed adducts. The exact origin of these adducts could not be accurately determined due to the multitude of possible etiologies. The capacity of the species to form genotoxin induced adducts was confirmed by a series of acute in vitro and in vivo laboratory exposures to 2-aminoanthracene, 2-aminogluorene, 2-acetylaminofluorene, benzo[a]pyrene and 4-nitroquinoline 1-oxide. These studies indicated that hepatopancreas adducts, in Mytilus spp., could be used as dosimeters of acute exposures to selected genotoxins. Further studies indicated that there was a delay in the maximum levels of adduct formation in Mytilus spp., following acute genotoxin exposure and that the species possessed the capacity to repair bulky genotoxin induced adducts. The persistence of these genotoxin induced adducts suggested however, that they could provide an indication of long-term exposure to selected genotoxic compounds. The standard 32P-postlabelling assay was inappropriate for the detection of adducts induced in Mytilus spp. following chronic exposures to either single, or complex mixtures, of genotoxins. In certain circumstances, the presence of pre-existing 'background' adducts compromised the analysis of genotoxin induced adducts. The application of the RSM assay was limited due to the lack of suitable DNA target sequences in Mytilus spp. The optimisation of available sequences together with recent advances in the assay's sensitivity and reliability, suggests that the assay has the potential for the analysis of the long-term effects of genotoxin exposure in Mytilus spp.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available