The Periplasmic Trehalase Affects Type 1 Fimbria Production and Virulence of Extraintestinal Pathogenic Escherichia coli Strain MT78.

Extraintestinal pathogenic Escherichia coli (ExPEC) is responsible for various infections outside the gastrointestinal tract in humans and other animals. ExPEC strain MT78 is invasive to various nonphagocytic cells and highly virulent in vivo To identify genes required for invasion of nonphagocytic cells by this strain, we applied signature-tagged mutagenesis to generate a library of mutants and tested them for invasion of avian fibroblasts. Mutants showing reduced cellular invasion included those with insertions in the fim operon, encoding type 1 fimbriae. Another attenuated mutant showed a disruption in the treA gene, which encodes a periplasmic trehalase. The substrate of TreA, trehalose, can be metabolized and used as a carbon source or can serve as an osmoprotectant under conditions of osmotic stress in E. coli K-12. We generated and characterized mutant MT78ΔtreA In contrast to the wild type, MT78ΔtreA was able to grow under osmotic stress caused by 0.6 M urea but not in minimal M9 medium with trehalose as the only carbon source. It presented decreased association and invasion of avian fibroblasts, decreased yeast agglutination titer, and impaired type 1 fimbria production. In a murine model of urinary tract infection, MT78ΔtreA was less able to colonize the bladder. All phenotypes were rescued in the complemented mutant. Our results show that the treA gene is needed for optimal production of type 1 fimbriae in ExPEC strain MT78 and that loss of treA significantly reduces its cell invasion capacity and colonization of the bladder in a murine model of urinary tract infection.

Enhancing in situ biodegradation in groundwater using pump and treat remediation: a proof of concept and modelling analysis of controlling variables.

A remediation approach which uses pump and treatment (PAT) to enhance the biodegradation of organic contaminants by increasing dispersive mixing between plumes and groundwater was evaluated for a phenol-contaminated aquifer, using a reactive transport model which simulates kinetic reactions between an electron donor (ED) in the plume and electron acceptor (EA) in the groundwater. The influence of system design and operation was examined in six modelling scenarios. Injection or extraction of groundwater increases biodegradation above no action and the location, pumping rate, and distance between well(s) are important variables which influence biodegradation. An increase in pumping rate, distance of the wells from the plume centreline, and changing the flow direction increase dispersive mixing between the plume and groundwater. This increases plume spreading and the plume fringe interface, providing a greater flux of dissolved EAs for biodegradation. In general, injection of groundwater containing natural EAs enhances biodegradation more than extraction. The enhancement of biodegradation is sensitive to the relative fluxes of ED and EA, as controlled by the arrangement of the wells. In the best performing scenario, biodegradation was enhanced by 128%, compared with no action.