Abstract: This study was undertaken to determine the prevalence of Salmonella sp. In human, animals and food items within Khartoum State, Sudan. Samples for analysis were collected from raw and cooked food items, fish, chlorinated drinking water, domestic livestock meat and poultry meat, livestock feces and human fecal samples for restaurant workers. Salmonella isolation and identification was performed according to standard previously described methods and the recovered isolates were serotyped and phagetyped at the Public Health Agency of Canada, Office International des Epizooties (OIE') Reference Laboratory for Salmonellosis. A total of 92 Salmonella enterica subspecies enterica isolates, representing 30 different serovars were recovered from 9.2% of the samples examined. Ninteen of the recovered Salmonella serovars were reported for the first time in Sudan. The latest study on Salmonellosis in Sudan dates to 1962 when Salmonella sp. were investigated only in animals. We examined various sources for the presence of Salmonella sp. therefore, this study provides a baseline study for comparison with any future Salmonella surveillance and epidemiological studies in Khartoum State, Sudan.