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Effective techniques for assessing soil environmental pollution are required to develop protective policy. Chemical methods have been traditionally used to determine total concentration of pollutants and biologically linked measurements have been used to assess the bioavailable fraction of pollutants. Bioluminescence-based microbial bioassays have been shown to respond to the bioavailable fractions. Growth and bioluminescence of lux-marked E. coli HB101 and P. fluorescens 10586r were characterised and optimised for freeze-drying culture. Freeze-drying cultures have been used effectively because of their ease of use, rapid assay response and sensitivity to a wide range of pollutants. An assessment of Zn and Cd amended soil was investigated. Two different techniques (centrifugation and Rhizon sampler) were used to obtain the interstitial pore water of soils. The concentrations of Zn and Cd were significantly higher in the soil solution extracted using the centrifugation technique compared to the Rhizon sampler technique. The biosensors responded to the free metal concentrations in the soil solution. An assessment of the toxicity of 2,4-dichlorophenol, 2,4,6-trichlorophenol and pentachlorophenol, individually and in combinations, was tested in deionised water (pH 5.5), soil solutions, and soils using lux-marked E. coli HB101 and P. fluorescens 10586r. Toxicity interaction responses of the mixture chlorophenols were predicted using a model. Synergistic interactions were observed for the response of P. fluorescens 10586r pUCD60-7 to all combinations of chlorophenol tested, while the response of E. coli HB101 pUCD607 varied with the matrix solutions tested. Bioavailability of naphthalene was studied using cyclodextrin-based extractions caused to the luminescence response of Pseudomonas fluorescens KH44 pUTK21. Increasing the concentrations of beta-cyclodextrin (b-CD) and hydroxylpropyl-b-cyclodextrin (HPBC) in the extract solutions increased the apparent concentration of naphthalene in the soil solutions. The luminescence response of P. fluorescens HK44 was associated with bioavailable of naphthalene.

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.

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.