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Biofilms are complex, sessile microbial communities that are problematic in clinical settings due to their association with survival and pathogenicity of bacteria. The biofilm formation supporting conditions for zoonotic serovars of Salmonella and avian pathogenic E. coli (APEC) from poultry have not been well studied yet. Clinical isolates of zoonotic Salmonella and APEC from poultry were evaluated for biofilm formation in four media at 37°C and 40°C after incubation of 48 and 72 hrs. The biofilms formed in 96 well plates were visualized and quantified with a new module of Aklides system using fluorescence microscope coupled with automated VideoScan Technology. After 72 hrs, brain heart infusion at 40°C and RappaportVassiliadis Soya broth at 37°C were found most suitable for APEC and Salmonella biofilm formations respectively. The new information will be useful for further biofilm associated studies particularly for evaluation of antibiofilm compounds and contribute in infection control. [ABSTRACT FROM AUTHOR]