Pneumonic pasteurellosis mainly caused by bacterial species of Mannheimia, Pasteurella, and Bibersteinia causes a significant financial loss to the sheep production sector through reduced productivity and high mortality. There is a dearth of information on the major agents involved in the disease in the Amhara region, Ethiopia. Therefore, the aim of this study was to isolate and molecularly confirm Mannheimia, Pasteurella, and Bibersteinia from nasal swabs of sheep suspected of pneumonic pasteurellosis in selected areas of the Amhara region. Isolation and phenotypic characterization were performed using microbiological and biochemical testing according to standard methods. Molecular confirmation of isolates was done through amplification of virulence associated genes, PHSAA and Rpt2, of Mannheimia hemolyticausing multiplex PCR. Accordingly, 46 out of 141 (32.62%) samples were presumably identified as M. hemolytica with no Pasteurella multocida and Bibersteinia trehalosi. Seven (n=7) out of the 46 isolates tested positive for either of the two virulence genes. The finding of this study is indicative that M. hemolytica is the main bacteria linked with pneumonic pasteurellosis in the study area which suggests the need to develop a polyvalent vaccine including strains of M. hemolytica or its antigenic determinants. However, the role of other bacterial, viral, and parasitic agents in the cases investigated should also be considered. © 2022 Akane et al.