Background: Salmonellosis is considered one of the most important infectious diseases in the developing countries where there is a significant lack of clean water supplies and poor sanitation in different areas helping the spread of the bacteria. It has a mortality rate of about 2–3% so, is considered a global health burden all over the world. Multidrug-resistant (MDR) strains of salmonella have been emerged making the inexpensive and readily available antibiotics including ampicillin, chloramphenicol, trimethoprim-sulfamethoxazole and streptomycin are frequently ineffective. Although resistance to ciprofloxacin, a second-generation fluoroquinolone, is increasing, it is still recommended as a first line therapy for almost all age groups. Objectives: Our study determines ciprofloxacin resistant salmonella strains in patients attending to Beni-Suef University Hospital and Beni-Suef Fever Hospital at Beni-Suef City. Also, this study assesses the capability of these strains to synthesize biofilm. Methodology: A total of 100 patients attended to Beni-Suef University Hospital and Beni-Suef Fever Hospital from December 2022 to March 2023 suspected clinically with salmonellosis were included. Stool samples were taken in suitable transport containers to Medical Microbiology and Immunology Laboratory Faculty of Medicine Beni- Suef University for further processing and identification. Cultures were done on MacConkey's media to detect lactose non fermenting strains and then on salmonella shigella SS agar to identify salmonella strains. Salmonella isolates are further identified by different biochemical tests. Minimum inhibitory concentration (MIC) for ciprofloxacin to the isolated salmonella strains using broth dilution method was determined. Ciprofloxacin resistant salmonella isolates were tested for the presence of biofilm by quantitative microtiter plates. Results: Over a period of five months, about 100 patients attended to Beni-Suef University Hospital and Beni-Suef Fever Hospital with a history and clinical symptoms of salmonella infection were involved in the study. The study included (36 males, 46 females) (54 child & 45 adults) (70 from rural while 30 from urban areas). Stool culture on MacConkey's and SS agar and typing of salmonella strains using API were done. MIC was determined for ciprofloxacin by broth dilution method. Biofilm evaluation of ciprofloxacin resistant salmonella isolates was determined using quantitative microtiter plates. Conclusion: We detected a high prevalence of salmonellosis, so continuous good preventive hygienic measures are needed. Biofilm production by salmonella strains made it more resistant to ciprofloxacin. These bacteria can form biofilm that aggravate the therapeutic problems in human and veterinary medicine, so continuous detection and evaluation is essential.