Abstract : Objective: The aim of this study, was detection of brucellosis in ewes by using Rose Bengal test (RBT), culture and polymerase chain reaction (PCR) technique as well as investigate histopathological changes in uterine tissue in ewes infected by Brucella melitensis. Methods: the study was conducted by using RBT, culture and PCR on the blood and uterine tissue, samples were taken from 139 of brucellosis suspected ewes, the study begging from November 2020 to July 2021 in Kerbala and Babylon provinces. Results: Serological, cultural and molecular examinations of samples of suspected infected ewes recorded positive cases with the observation of histopathological changes of the uterus. 139 ewes were examined and show different sings like abortion, fever, depression, loss of weight and ewes that gave birth to weak lambs, the result recorded by the PCR was 37 (26.61%) positive cases, less than that recorded by the Rose Bengal test, which was 41 (29.49%), while the culture showed 35 (25.17%) positive cases of brucellosis. The microscopical analysis of uterine samples that collected from infected ewes by Brucella melitensis revealed marked histopathological changes. The marked histopathological changes were showed a different severity including infiltrations of mononuclear cell, fibrosis, necrosis, hemorrhage, calcification of tissue and severe congested blood vessels. Conclusion: This study revealed prevalence of brucella melitensis in sheep. The molecular technique and culture showed the most accurate test for detection of brucellosis in animals, brucella melitensis infection was detected by using PCR test which considered important to discover the brucellosis infection after use the fast routine screening of herds like RBT. The slaughtered ewes that infected by brucella melitensis showed various histopathological changes in the uterine tissue that effect on the health status of the ewe and cause abortion. Key words: RBT, PCR, molecular tests, histopathological, Brucella melitensis, ewes.