Anthrax is a zoonotic disease caused by Bacillus anthracis (B. anthracis), a grampositive,rnnon-motile, and spore-forming bacterium. Animal anthrax is an important andrnzoonotic disease in Ethiopia and only few of those cases were isolated by laboratory.rnOutbreak investigation of anthrax suspected cases was conducted from November 2018rnto June 2019 in south Gonder, Illubabor, Wolayita, Gurage Zone and Zone 4 of Afarrnregion of Ethiopia, where five different outbreaks in different animal species werernreported. Purposive sampling method was used in suspected outbreak cases in domesticrnanimals (cattle and goat) and wild animals such as hippopotamus. Anthrax suspectedrnsamples were tested under appropriate biosafety and containment. A total of 63 differentrnsamples like nasal swab, rectal swab, tissue, and swab from old carcasses andrnenvironmental samples were collected, labeled, packed and transported to NAHDIC,rnSebeta for further processing. Isolations of B. anthracis was conducted by culturing onrnblood agar, colony characterization, Gram staining, spore and capsule staining andrnbiochemical tests. The molecular detection was conducted by real-time polymerase chainrnreaction. 9.8% (6/61) of B. anthracis was identified from dried meat (Quwanta) samplesrnfrom outbreak cases in South Gondor, Farta District and the Bacillus isolates were nonhemolytic,rnnon-motile and susceptible to penicillin, which is typical of B. anthracis. Thernidentified colonies were further confirmed by real-time PCR, where B. anthracis virulentrnplasmid markers namely protective antigen (on plasmid pXO1) and capsule (on plasmidrnpXO2) were detected. This is the first report of PCR confirmed anthrax outbreak casernfrom animals in Ethiopia. Anthrax still remains endemic in the country and causesrndiseases in livestock. A multi-sectoral collaborative approach and capacity buildings arernessential for successful prevention and control of the disease and joint outbreakrninvestigation, proper and timely animal immunization.