The objective of this research was to determine the potential of two anti-atrazine single-chain antibodies (scAbs) to detect very dilute atrazine concentration, specifically in the nanomolar and sub-nanomolar ranges. Analysis of environmental pesticide contamination is routinely done by high performance liquid chromatography (HPLC), gas chromatography (GC) and mass spectrometry (MS). However, since the mid eighties there have been an increasing number of reports describing the use immunoassays to detect environmental pollutants. Although immunoanalysis using whole antibodies has been shown to be almost as effective as GC/MS (Thurman et al., 1990) the technology is still not widely used for pesticide analysis. This thesis will describe the increased sensitivity of single-chain antibody fragments, compared to whole antibodies, to detect low concentrations of atrazine and related triazines. The effect of monomeric and dimeric scAb conformations on the ability of enzyme linked immunosorbent assays (ELISAs) to detect atrazine and related triazines is investigated. The thermal and chemical stability of the scAbs, and a modified scAb containing an interchain disulphide bond, are compared with the parent monoclonal antibodies (MAbs) to determine their suitability in an environmental ELISA assay system. The expressions in Escherichia coli (E. coli) of two different anti-atrazine scAbs and related antibody fragments are studied, and an optimised expression protocol for these antibody fragments obtained. Three potentially toxic amino acid residues identified in one of the variable heavy (VH) domains (Knappik & Pluckthun, 1995) are mutated to less toxic amino acids, and their effects on antibody fragment expression observed.

A novel method for the bioremediation of phenanthrene using the fungus Penicillium frequentans was utilised to remove phenanthrene (200 mg kg\(^{-1}\)) from soil containing both metals and phenanthrene, over 29 days. Bioremediation of phenanthrene and its effects on trace metal behaviour has been investigated. Metal behaviour studied includes metal speciation and the kinetics of exchange between solution and solid phase and plant uptake of the more labile and mobile, and potentially more bioavailable metal species. Phenanthrene removal by P. frequentans was optimised in terms of both soil water and nutrient composition. Slightly lower removal rates were obtained using P. frequentans alone (73%) and plants alone (67%). However, the highest phenanthrene removal (77%) was obtained using both fungus and plant. Assessment of the metal behaviour before and after phenanthrene biodegradation showed that the removal of phenanthrene by either fungal or mixed fungal and native microflora resulted in an increased flux of metal from solid to solution, an increased pool of potentially bioavailable and toxic metal species and increased plant uptake to both Echinochloa polystachia and Triticum aestivum, by factors of 4-13. In the presence of plants alone, metal mobilisation and uptake increased by smaller factors. In some cases, there was no increase in metal mobilisation and a maximal increase of 2 was found in Ni and Pb. These results highlight the impact of bioremediation process on metal behaviour. In addition, it is suggested that phytoremediation and not bioaugmentation using P. frequentans is the best overall option to obtain a considerable phenanthrene removal, reducing the increased pool of potentially bioavailable and toxic metal species

The adsorption of sulphate has been studied using alpine and sub-alpine podzolic soils from the Allt a'Mharcaidh catchment, Cairngorms, Scotland, U.K. A detailed investigation of sulphur pools and mechanisms of sulphur retention has revealed that incoming sulphate ion, both marine and anthropogenic, is being retained by a range of adsorption mechanisms. Analysis of the soils has indicated that the sulphur content of the soils is dominated by organic forms of sulphur. Selective chemical fractionation techniques have identified an accumulation of sesquioxides, dominated by amorphous aluminium, in the basal horizons of all the soil profiles. Adsorption of sulphate has been shown to be mainly governed by the quantity and variable charge character of this sesquioxide phase. Fractionation has also suggested that the sesquioxides in the basal horizons of the soils are dominated by a combination of gibbsitic and imogolitic materials, the latter being present as a result of podzolization processes. The generation of sulphate adsorption isotherms has revealed that the organic horizons of the catchment soils show little or no sulphate adsorption capacity, due to a small sesquioxide content. In contrast, the mineral horizons are all capable of adsorbing sulphate, and in most cases this adsorption can be modelled to the Langmuir adsorption equation. The adsorption capacity of particle size fractions of the soils has revealed that even soil particles > 1 mm show an ability to retain sulphate, possibly by physical entrapment of soil solution in mineral pores or amorphous silica gel coatings. The accompanying hydroxyl release that is generally assumed to accompany adsorption of sulphate in the mineral soils was not evident, and unless a ligand exchange reaction was masked by other chemical reactions involving total proton balance, sulphate adsorption in the catchment soils is not due to a specific or 'low affinity' specific mechanism.

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