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Lencho Megersa Marami,1 Getachew Mulatu Dilba,1 Dagmawit Atalel Babele,1 Edilu Jorga Sarba,2 Askale Gizaw,1 Wakuma Mitiku Bune,1 Morka Dandecha Bayu,1 Petros Admasu,2 Abraham Mekbeb,2 Miressa Tadesse,3 Kebede Abdisa,2 Dejene Bayisa4 1Ambo University, College of Agriculture and Veterinary Science, Department of Veterinary Laboratory Technology, Ambo, Oromia, Ethiopia; 2Ambo University, College of Agriculture and Veterinary Science, Department of Veterinary Science, Ambo, Oromia, Ethiopia; 3Ambo University, College of Computational and Natural Science, Department of Chemistry, Ambo, Oromia, Ethiopia; 4Jeldu Woreda, Livestock and Fishery Development Office, Gojo, Oromia, EthiopiaCorrespondence: Getachew Mulatu DilbaAmbo University, College of Agriculture and Veterinary Science, Department of Veterinary Laboratory Technology, P.O. Box 19, Ambo, Oromia, EthiopiaTel +251-912717217Email getachew.mulatu@ambou.edu.etBackground: Although traditional healers in Ethiopia have a long history of using medicinal plants to treat diseases in animals and humans, studies on the antibacterial activities and potential bioactive ingredients of most medicinal plants have been insufficient. Therefore, this study aimed to evaluate the in-vitro antibacterial activities and to screen phytochemical constituents of selected medicinal plants against reference bacterial strains.Methods: The fresh and healthy roots of Echinops amplexicaulis, fruits of Ruta chalepensis, and leaves of Salix subserrata were collected from West Shewa Zone, Ethiopia. Agar well diffusion and agar dilution methods were used to evaluate antibacterial activities and minimum inhibitory concentrations (MIC). All the crude plant extracts were tested against Staphylococcus aureus, Streptococcus pneumoniae, Escherichia coli and Pseudomonas aeruginosa at concentrations of 100, 50, and 25 mg/mL in each triplet (3x). MIC of crude extracts ranging from 1.5625 to 12.50 mg/mL was applied to all bacterial strains. The positive control