Human cases of salmonellosis are often linked with the consumption of poultry-based products contaminated with pathogenic serotypes of Salmonella. Chickens colonised with Salmonella often do not show clinical disease; therefore, it is not known what dose is field relevant for establishing gut colonisation. Moreover, the role dust has in Salmonella transmission on farm is not well characterised. In this study, the role of route of inoculation and the threshold of Salmonella Typhimurium infection in chicks using dust as a carrier was investigated. In a pilot study, layer chicks were infected with Salmonella Typhimurium prepared in dust (106 CFU/g) and administered either by sprinkling over birds or via oral administration. In a subsequent dose response experiment, layer chicks were infected by sprinkling method with Salmonella Typhimurium using 102, 104 and 106 CFU/g of dust. In both studies, Salmonella was detected in cloacal swabs on day 1, 3 and 5 post-infection (p.i.) and was quantified from liver, spleen, lungs and caecal tissues 1 week p.i., with a minimum dose of 104 CFU required. Furthermore, chicks inoculated through sprinkling had significantly higher Salmonella loads in liver and spleen at week 1 p.i when compared to oral gavage. Salmonella count data obtained 1 week p.i. showed that 102 CFU treatment did not result in systemic infection as no bacteria were detected in liver, spleen, lungs, or caecal samples. Irrespective of the infection dose (104 or 106 CFU/g of dust), Salmonella colonised the gut and invaded into internal organs at a similar level. These data show that dust sprinkling is more effective at allowing colonisation of chicks with Salmonella than oral gavage. As dust is likely to play a role in horizontal transmission in chicken sheds. This study establishes a foundation model appropriate for laboratory based studies of field challenge and will be essential for future assessment of vaccination strategies for the control of wild-type Salmonella in poultry production.