Environmentally persistent Salmonella in the pet food factory environment has been described, with biofilm formation suggested as a candidate mechanism contributing to their persistence. In this study the ability of a panel of Salmonella isolates from factory, clinical, and veterinary sources was investigated for their ability to form biofilms at 24 and 48 hours. The effect of nutrient availability and incubation time on biofilm formation was investigated using full strength and diluted 1/20 TSB media at 37°C, 25°C, 15°C, and 10°C. Results highlighted that all the Salmonella isolates were able to form biofilms in both nutrient conditions and this was highly correlated with temperature. At 25°C, biofilm formation was enhanced in diluted 1/20 TSB and increased incubation time (48h) (p= <0.001). However, this was not observed at 10°C, 15°C, or 37°C. None of the factory isolates demonstrated enhanced biofilm formation in comparison to serotype-matched isolates from veterinary and clinical sources. Salmonella enterica Senftenberg 775W was the strongest biofilm former at 15°C, 25°C, and 37°C in all the conditions tested (p=<0.05). Biofilm formation is an important mechanism of environmental persistence in the food manufacturing environment; however, there is no evidence of an enhanced biofilm-producing phenotype in factory persistent strains.