Brucellosis in Egypt is still one of the most serious problems facing animal production. This study aimed at detection of brucellosis in animals and man in New Valley Governorate. Serum samples of cattle, sheep, goats and camels were examined by Rose Bengal plate test, buffered acidified plate antigen test, tube agglutination test and Rivanol test. Serological examination of blood samples revealed that the prevalence of brucellosis among cattle, sheep and goat by using RBPT, BAPAT, TAT and RT was (10.56%, 7.78%, 7.22% and 7.22%),(3.89%, 2.78%, 2.22% and 2.22%), (7%, 7%, 6% and 6%). Brucellosis could not be detected in El-Rashda and El-Owinat. The infection was higher in female than males. The infection rate of brucellosis was studied in different ages. Brucellosis could not be detected in camels. The results of human screening by using RBPT, TAT & RT revealed a prevalence of brucellosis with rates of 8.18%, 7.27% and 5.45%, respectively. The majority of human cases occurred in butchers, veterinary attendants, housewives, veterinarians, children and farmers. The prevalence was higher in females (9.09%, 9.09% & 5.45%) than males (7.27%, 5.45% & 5.45%). The human age–wise was found maximum at age group 21-40 years (11.11%, 9.52% & 6.35%) followed by age 6-20 years (8.33%, 8.33% & 8.33%) then age 41-60 years (5.56%, 5.56% & 5,56%) and could not be detected at age group >60years. All human cases were detected in rural areas.