A range of novel substrates for the detection of bacterial hydrolyases have been examined in both liquid and solid media. The potential inhibitory effect of these substrates on both Gram-positive and Gram-negative organisms has been evaluated. In addition a range of alternative substrates possessing an appropriate chemical configuration have also been evaluated as substrates in this thesis. It has been demonstrated that substrates based on 4-aminophenol, particularly the dichloro derivative produce intensely coloured reaction products upon hydrolysis and subsequent coupling with 1-naphthol. A derivative of 1-naphthol, 3,5-dihydroxy-2-naphthoic acid, was shown to react favourably with a number of released core compounds, producing a wide variety of coloured complexes. In comparison with o-nitropheny1-13-D-galactoside (ONPG), the novel substrates for the detection of P-galactosidase activity showed wide differences in K. and V. values. Overall these substrates performed well in liquid media, especially the dichloro derivatives for detection of a range of bacterial hydrolyases. The substrates have also been evaluated in solid media and have been shown to produce intensely coloured reaction products. The fact that these substrates offer themselves to the production of novel dual substrate systems indicates their potential diagnostic applications. Use of L-alanyl-DEPPD in combination with 1-naphthyl-J3-D-galactoside has been shown to be of diagnostic potential for the detection of 13-galactosidase activity in Gram-negative organisms. In addition, a combination of substrates, L-prolyl-4-amino-2,6-dichlorophenol and 1-naphthyl-O-D¬glucoside has also been shown to have potential for the production of a medium specifically for the isolation of Serratia sp. This dual substrate system also has applications for the production of media for the specific isolation and detection of numerous clinically important pathogens e.g Enterococcus sp, C.perfringens and Yersinia enterocolitica.

The detection and control of microorganisms such as bacteria is important in a wide range of industries and clinical settings. Detection, binding and removal of such pathogenic contaminants can be achieved through judicious consideration of the targets which are available at or in the bacterial cell. Polymers have the ability to present a number of binding ligands for cell targeting on one macromolecule and so avidity of interaction can be greatly increased. The goal of the project was to test whether polymers generated with bacteria in situ would have their composition significantly altered to determine if a templating process was occurring. It was also anticipated that the templated polymers would have better re-binding properties than those produced in the absence of bacteria. A series of chemical functionalities were analysed for their binding properties to bacteria. The functionalities were chosen with consideration to the cell surface characteristics. Further to identification of the most binding and least binding functionalities the polymers were tested for their cytotoxicity against bacteria and human epithelial cells. Concentration ranges were determined which could facilitate bacterial binding and templating yet minimise the lethality of the processes. Templated polymers of the bacteria were generated using a novel method of atom transfer radical polymerisation (ATRP) which we have termed bacterial activated atom transfer radical polymerisation (b-ATRP). This polymerisation method has maximised the potential for templating processes to occur during the polymerisation. Templated polymers differed in both their composition and their binding behaviour to non-templated polymers. The bacterial organic reduction process has also been demonstrated to have greater scope for use within the organic chemistry field as demonstrated by the use of this system to enable in "click-chemistry" via the reduction of copper.