This research explored the possible application of pressurised carbon dioxide (P CO2), a promising non-thermal sterilisation technique, for the treatment of sewage sludge (SS) before anaerobic digestion to inactivate pathogenic microorganisms. Escherichia coli was selected as the test organism and was isolated from SS and maintained in pure culture. The growth curve of the isolated strain was determined by measuring the optical density (OD) in liquid culture medium and relating this information to the spread plate count so that a culture of known cell density could be grown for optimisation experiments. Inactivation of E. coli was enhanced by increase in pressure (1,500, 2,000 and 2,800 kPa) and treatment time (from 0.75 to 24 h). A short exposure time at high pressure was sufficient to provide a degree of inactivation which could also be achieved by longer exposure at lower pressure. Complete inactivation (8 log10 reduction) was possible at all three pressures. scanning (SEM) and transmission (TEM) electron microscopy studies of E. coli treated with P CO2 revealed that the cell walls were ruptured, and the cytoplasm was unevenly distributed and had lost its density, indicating the possible leakage of intracellular substances.

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