This study aimed to evaluate the knowledge and attitudes of community pharmacists (CPs) on vaccination and assess the barriers and willingness to implement community pharmacy-based vaccination services (CPBVS) in Ethiopia. An online cross-sectional study was conducted on 423 CPs in Ethiopia, and questionnaires were distributed to CPs through the Ethiopian Pharmaceutical Association telegram group and e-mail invitations. Data were analyzed using the Statistical Package for Social Sciences (SPSS) version 27. Most CPs (92%) had good knowledge of vaccination, and 43.5% strongly agreed that the population’s immunization rates would rise if CPs were authorized to provide vaccinations. The overall mean attitude score (±SD) toward vaccination was 35.95 (±4.11) out of a total score of 45, with 187 (44.2%) scoring below the mean. The most common barriers were lack of authorization (94.1%), costs and time associated with professional development and training (71.4%), time requirements for professional development (70%), and insufficient staff or resources for implementation (70%). Two hundred thirty CPs (54.4%) expressed a willingness to implement CPBVS. Educational qualifications were significantly associated with knowledge of CPs regarding vaccination. Those with inadequate knowledge had about 2.5 times (AOR = 2.51, 95% CI: 1.19, 5.31,
p
= .016) a poorer attitude toward vaccination services compared with those with adequate knowledge. Those study participants who had a good attitude toward vaccination services were nearly seven (AOR = 6.80, 95% CI: 4.36–10.59,
p
= .0001) times more willing to provide CPBVS when compared with their counterparts. Implementing CPBVS in Ethiopia requires overcoming barriers and providing professional development opportunities.