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This study involved cross-sectional serological and questionnaire-based surveys to investigate the sero-epidemiology of foot and mouth disease (FMD) in domestic ruminants, and farmers' knowledge and practices about the disease in the Amhara region of Ethiopia. A multistage cluster sampling was carried out to select domestic ruminants for serological sampling and for the interview with farmers. A total of 1,672 sera samples were collected and tested using a 3ABC-Enzyme Linked Immunosorbent Assay, and 170 farmers were interviewed. An overall FMD apparent seroprevalence of 11.48% (95% CI: 7.52–17.14%) was recorded in the domestic ruminants. The overall true prevalence was 12.04%. The seroprevalence of FMD was higher in cattle (14.37%) than in goats (7.10%) and sheep (7.07%). The age stratified seroprevalence in the districts showed that 66.67% of the districts studied experienced a FMD outbreak within the preceding year of the study time. A mixed effect logistic regression analysis revealed that agroecology, the production system and the age of the animal was significantly associated with FMD seropositivity in cattle (P < 0.05). A statistically significant (P < 0.05) positive correlation (r = 0.93) was observed between cattle and small ruminant FMD seroprevalences. About 82% of the farmers interviewed knew of FMD and 85% of them had experienced the disease in their own herds before. The farmers mostly employ traditional means to control FMD. In conclusion, the findings of the study indicated that FMD is a prevalent disease in the Amhara region with more importance in the intensive production systems and the lowlands of the region. High correlation in seroprevalence between small and large ruminants indicated a possible cross transmission between these species. Therefore, small ruminants should not be overlooked in FMD control. Farmers in the region have a good level of knowledge about the disease; however, currently they heavily rely on traditional practices primarily focused on treating wounds of infected animals. This calls for extension work on available effective preventive measures of the disease, such as vaccination and movement restriction.