In the current study, a total of 50 sheep, 23 goat and 60 human blood serum samples were collected for serological tests. A total of 30 (22 ewes and 8 does) milk samples, 43(25 ewes and 18 does) vaginal swabs, 14 preputial swabs from rams and 60 human citrated plasma were collected for bacteriological and molecular studies. All examined sheep and goat were non-vaccinated. Ewes and does had history of abortion, while rams had no clinical signs. Samples were collected from different localities at Minia governorate. Serological results using buffered acidified plate antigen test (BAPAT), rose Bengal test (RBT), rivanol test (RT), tube agglutination test (TAT), and indirect enzyme linked immunosorbent assay (iELISA) gave 32(64%), 30(60%), 26(52%), 31 (62%), and 28 (56%) in sheep respectively, where in goat revealed 23(100%), 23(100%), 22(95.7%), 23(100%), and 22(95.7%) respectively. While human serum samples were 33(55%), 29(48.3%), 33(55%), 27(45%), and 23(38.3%) respectively. The bacteriological results revealed 35 isolates [10(45.5%), 4(50%), 4(16%), 10(55.6%), 2(14.3%), and 5(8.3%)] from ewes' milk samples, does' milk samples, ewes' vaginal swabs, does' vaginal swabs, rams' preputial swabs and human citrated plasma respectively. All 35 isolates were molecularly identified using polymerase chain reaction (PCR) as B. melitensis biovar3. Conclusively, brucellosis was endemic at Minia governorate with a high level especially in goat. Rams act as chronic carriers leading to public health hazards. B. melitensis is the prevalent serotype in Egypt. More effective preventive and control measures should be applied