Abstract : Brucellosis is a frequent infectious zoonosis over the world, intimidating both veterinary and human public health. The misuse and overuse of antibiotics in Iraq in the veterinary field is associated with the high rate of antibiotic resistance that observed in some zoonotic bacteria directing to cause serious infections in human. The point of this study; is to determine the occurrence of extended-spectrum beta-lactamases (ESBLs) -producing Brucella abortus in the blood of cattle. One hundred blood samples were obtained from cattle in a number of slaughterhouses in different areas of Baghdad. Equal to 10 ml of blood samples were collected from the tail vein of the animals. Blood was divided into two parts; the first part (7ml) in Trypticase soy broth, incubated at 37 °C with 5–10% CO2 for 2-3 days. Later, Bacterial DNA was extracted and a specific set of primers were used for the detection of β-lactamases genes in Brucella abortus isolated using multiplex-PCR technique. of 100 blood samples used in this study 26 isolates (26%) were confirmed to be Brucella abortus species by VITEK-2 System. The ESBLs genes; blaTEM gene was detected in 9 of 26 (34.6%) isolates, followed by blaCTX-M gene in 7(26.9%) isolates, blaAMPC gene in 6 (23.0%) isolates, and blaSHV gene in 4 (15.3%) isolates. This study indicated the high rate of ESBLs- producer Brucella abortus among cattle. Thus, indicating the potential of animals as a source of the ESBLs-producer bacteria that transmitted to human via the food chain or other recreational activities.