Background: Brucellosis is an infectious disease caused by bacterial of the genus Brucella. Although several investigations detected Brucella antibodies in many animal species, low attention has been paid for specific identification of different Brucella species in animals of Sub-Saharan African counties. It is in this light that this study was designed to identify Brucella abortus and Brucellla melitensis in cattle and sheep of several villages of southern Cameroon in order to improve our epidemiological knowledge on brucellosis in central AfricaMethods: Blood samples were collected in 597 cattle and 315 sheep of 15 villages of two regions of Cameroon. DNA was extracted from blood samples and primers amplifying bcsp31 gene enabled the identification of Brucella spp infections in domestic animals. Amongst animals found with Brucella spp infections, specific primes were used to identify B. abortus and B. melitensis respectively. Results: From 912 animals analyzed in this study, 159 (17%) were found with Brucella infections. This gives an infection rate of 20.93% (125/597) in cattle and 10.79% (34/315) in sheep respectively. Of the 159 animals harboring Brucella infections, 118 (12.93%) were infected by B. abortus and 29 (3.17%) by B. melitensis. The Brucella infection rate (20.56%) was high in animals of the Noun division compared to those (12.35%) of Yoko. Between animal species, significant differences were observed in infection rates of both abortus and B. melitensis. The difference in the B. abortus infection rates remains significant between Yoko and Noun, and especially between villages of the Noun division. Conclusion: This study revealed for the first time B. melitensis and B. abortus infections in cattle and sheep of the Noun and Yoko in the southern Cameroon. The identification of B. melitensis and B. abortus in animals suggests further investigations on human brucellosis. Results of this study highlight the need of developing and implementing control measures against brucellosis in Cameroon, especially in villages presenting high risk for Brucella infections.