From a collection of over 2800 Salmonella enterica subspecies Enterica serotype Typhimurium F98 Tn5-TC1 insertion mutants 14 were identified as expressing growth-non-suppressive phenotype under strict anaerobic conditions. Sequence analysis of regions flanking the Tn insertions revealed that most of the selected mutants were defective in genes contributing to the anaerobic fumarate uptake and generation (insertions in dcuA, dcuB and aspA), or to the anaerobic L-arginine utilisation pathway (insertions in STM4467 encoding a putative arginine deiminase, and in between speF encoding ornithine decarboxylase and kdpE coding a response regulator protein). Mutants defective in flagellum synthesis (flhA) were also identified. In contrast to the in vitro results, all the mutants colonised 1-day-old chicks efficiently and suppressed the super-infection of chicks by the parent strain. This clearly indicates that neither of the metabolic pathways mentioned above nor motility play essential roles in lower intestinal tract colonisation.