This study aimed to assess the prevalence of Salmonella spp. and Pseudomonas spp. in poultry farms across Assiut governorate, Egypt. A total of 600 environmental samples were collected from 11 farms between July 2016 and February 2017. Samples were analyzed using conventional culture methods, with presumptive colonies subjected to biochemical tests, followed by molecular identification via multiplex and conventional PCR. Of the collected samples, 129 (21.5%) tested positive for Salmonella spp., and 81 (13.5%) for Pseudomonas spp. Molecular analysis of 35 Salmonella isolates revealed 19 (54.29%) as Salmonella typhimurium and 2 (5.71%) as Salmonella enteritidis. Additionally, 4 (26.67%) of 15 Pseudomonas isolates were confirmed as Pseudomonas aeruginosa. The findings demonstrate significant bacterial contamination, underscoring the role of poultry farms as reservoirs for these pathogens. Implementing proper management practices and targeted interventions is essential to mitigate the risk of contamination in poultry environments.

Keywords: prevalence, poultry farms, Salmonella spp., Pseudomonas spp., environment, Assiut, Egypt