High_through_put_phenotyping_of_Salmonella

almonella enterica subsp. enterica can be divided into multiple serovars based on phenotypic (flagella, O-antigen, capsule) and biochemical properties, as well as now through genetic typing methods (MLST, WGS). These serotypes can be broadly further classified as host-restricted, host-adapted “specialists” or host-unrestricted “generalists” based upon their host range and common clinical manifestation of associated infections. As we have now sequenced thousands of Salmonella there is a push to move from genotyping to broad phenotyping. The aim is to link genotype to phenotypes, including the genetic mechanisms of host adaptation and disease pathology. Or collaborators (Peter Schierack & Rafal Kolenda) have developed an automated microscopy-based bacterial infection assay they have named VideoScan (1) for monitoring the ability of S. entrica isolates to interact with host cells. VideoScan is a relatively low resolution system but it can be performed at reasonable scale. Using this method, they have determined the cellular infection phenotypes for 127 Salmonella isolates covering five S. enterica serovars. This analysis includes their ability to infect three different cell lines (IPEC-J2, Caco-2 and CHIC8-E11). In this analysis they found striking differences between 'generalists' and 'specialist' isolates in terms of adherence and invasion, but also in their ability to grow in cell culture medium. We would like to perform a more detailed phenotypic analysis, including RNA-seq, on these isolates. Unfortunately, they have not been sequenced, limiting our ability to link genotype to phenotype. Thus, we will carry out whole genome sequencing of 30 of these Salmonella isolates representing a diverse range of infection phenotypes. Understanding the factors influencing infectivity and virulence will provide useful information for treatment and prevention of Salmonella infections. As part of this collaboration, a postdoctoral fellow will work at the Sanger for several months to develop the project and collect more phenotypic data. (1) A highly versatile microscope imaging technology platform for the multiplex real-time detection of biomolecules and autoimmune antibodies. Rödiger S, Schierack P, et al. Adv Biochem Eng Biotechnol. 2013;133:35-74.