Animals including the poultry and its by product are major sources of human salmonellosis as well as animals raised for food play an important role in transmission of antimicrobial resistant Salmonella strains to humans. This study was under-token to determine the prevalence of Salmonella spp. In human, animals, food items, and their antimicrobial resistance patterns in Najran region, Saudi Arabia. Samples for analysis were collected from human (n. = 532) and animals ((sheep n. = 330, camels n. = 44 and cattle n. = 59) faecal samples, Poultry by product (meat) (n. = 120) including egg (n. = 506), wild pigeons (n. = 43), Fish (n. = 85) and fresh she-camels milk (n. = 13). Salmonella isolation, identification and antibiotic sensitivity were performed according to standard previously described methods. The prevalence of Salmonella isolates in human, sheep, camel and cattle was (2.4%, 6.4%, 2.3% and 8.5) However, the prevalence of Salmonella isolates in fish, poultry meat, table eggs, fresh camel milk and wild pigeon was (0 %, 4.1%, 2.1%, 0%, 11.6%, and 2.7%).The prevalence of Typhoid and non-typhoid salmonellosis were 46.1% and 53.8 % respectively. Isolated Salmonella spp. were Salmonella subsp.1, S. arizonae subsp.3A, Salmonella subsp.5, Salmonella typhimurium, Salmonella typhi, Salmonella paratyphi A and Salmonella pullorum. (37.7%, 3.3%, 4.9%, 24.6%, 8.2%, 1.6% and 19.7%) Respectively. All isolated salmonella sp. were significantly higher sensitive (P<0.05) to Ciprofloxacin (5μg) (97.1 %). In conclusion, the serious implications associated with drug-resistant Salmonella species, a more deliberate use of antibiotics in both human medicine and animal industry is warranted. Furthermore, attentions must be taken to wild pigeons that played important role as source of Salmonella typhimurium infection to human, animals and domestic birds in Najran region, Saudi Arabia. Non-Saudi Resident workers should be have health certificate and periodically renewed specially Restaurant workers because they were a source of new isolates, serotypes and clones of Salmonella species in Saudi Arabia.