Bacteria-protozoa interactions are one of the oldest between prokaryotic and eukaryotic organisms. As such, their study offers a unique opportunity to understand the different relationships that have evolved between them, including pathogenesis, and how their interaction can affect some important processes, such as wastewater treatment. In the first part of the work described here, the grazing defence mechanisms employed by Pseudomonas aeruginosa against the surface grazer, Acanthamoeba castellanii, were investigated. P. aeruginosa cells from early logarithmic growth and stationary phase were found to use different defence strategies. The type-III secretion system (T3SS) was found to be responsible for cytotoxicity of early logarithmic growth cells against A. castellanii. Of the three exotoxins produced by P. aeruginosa PA99, the phospholipase ExoU was found to make the greatest contribution to bacterial toxicity against the amoebae. Interestingly, a PA99null mutant that does not produce any known exotoxins but synthesises a secretion apparatus, was also found to be toxic to the amoeba, suggesting that the T3SS was being used to translocate other unknown toxins. Quorum sensing regulated virulence factor production was found to be involved in the grazing defence response of stationary phase P. aeruginosa cells. A. castellanii was found to be most susceptible to hydrogen cyanide and elastase produced during late logarithmic and stationary phase. In the second part, a stable isotope probing method was developed to investigate carbon flow through bacteria-protozoa food webs in activated sludge. The method was subsequently used to track carbon from bicarbonate and acetate through bacteria-orotozoa food webs under ammonia oxidising and nitrate reducing conditions. It was found that the Peritrich ciliate Campanella umbellaria, dominated the acquisition of carbon from bacteria with access to CO2 under ammonia oxidising conditions. Thus it appears that some of these bacteria must live in the plankton, as C. umbellaria is a filter feeder. No specific protozoan groups were found to dominate carbon acquisition from bacteria with access to acetate, under nitrate reducing conditions, probably due to label dilution. Overall the results presented here showed how bacteria-protozoa interactions have shaped infectious processes in higher eukaryotes, and the dynamics of carbon flow in activated sludge.

Bacteria-protozoa interactions are one of the oldest between prokaryotic and eukaryotic organisms. As such, their study offers a unique opportunity to understand the different relationships that have evolved between them, including pathogenesis, and how their interaction can affect some important processes, such as wastewater treatment. In the first part of the work described here, the grazing defence mechanisms employed by Pseudomonas aeruginosa against the surface grazer, Acanthamoeba castellanii, were investigated. P. aeruginosa cells from early logarithmic growth and stationary phase were found to use different defence strategies. The type-III secretion system (T3SS) was found to be responsible for cytotoxicity of early logarithmic growth cells against A. castellanii. Of the three exotoxins produced by P. aeruginosa PA99, the phospholipase ExoU was found to make the greatest contribution to bacterial toxicity against the amoebae. Interestingly, a PA99null mutant that does not produce any known exotoxins but synthesises a secretion apparatus, was also found to be toxic to the amoeba, suggesting that the T3SS was being used to translocate other unknown toxins. Quorum sensing regulated virulence factor production was found to be involved in the grazing defence response of stationary phase P. aeruginosa cells. A. castellanii was found to be most susceptible to hydrogen cyanide and elastase produced during late logarithmic and stationary phase. In the second part, a stable isotope probing method was developed to investigate carbon flow through bacteria-protozoa food webs in activated sludge. The method was subsequently used to track carbon from bicarbonate and acetate through bacteria-orotozoa food webs under ammonia oxidising and nitrate reducing conditions. It was found that the Peritrich ciliate Campanella umbellaria, dominated the acquisition of carbon from bacteria with access to CO2 under ammonia oxidising conditions. Thus it appears that some of these bacteria must live in the plankton, as C. umbellaria is a filter feeder. No specific protozoan groups were found to dominate carbon acquisition from bacteria with access to acetate, under nitrate reducing conditions, probably due to label dilution. Overall the results presented here showed how bacteria-protozoa interactions have shaped infectious processes in higher eukaryotes, and the dynamics of carbon flow in activated sludge.