A cross-sectional study was carried out during 2022 to evaluate the current status of bovine brucellosis at Aswan governorate as a border governorate and where an extensive process of live animal importation occurs through Abu Simbel quarantine.  The present study estimated the seroprevalence of brucellosis among local and imported cows, identified the currently field circulating Brucella strains in Aswan governorate and described the most important risk factors associated with the dynamics of bovine brucellosis. A total of 932 cows including 145 imported cows and 787 local cows from different localities, 319 ewes and 581 does at Aswan governorate were sero-tested for brucellosis using Rose Bengal plate test (RBPT), and Buffered acidified antigen plate test (BAAPT) as screening tests and positive samples were confirmed by complement fixation test (CFT). The seroprevalence of brucellosis was estimated as 2 (1.4%) in imported cattle and 20 (2.5 %) in local cattle with overall prevalence of 22 (2.4%) in cattle, 4 (1.2%) ewes and 6 (1%) in does. Different clinical specimens including 10 cow's milk samples, 5 cow's aborted material and 5cow's manure samples were collected. Clinical samples were processed for bacterial isolation and PCR testing. A total eight Brucella melitensis biovar 3 and two Brucella abortus biovar1 isolates were recovered and identified on bacteriological and molecular bases. Brucella melitensis biovar 3 could be isolated from two manure samples of infected cows that suggests that manure could be a vehicle for indirect dissemination of infection in both animals and man and could be an important potential risk factor.