Fusarium wilt of cucumber caused by Fusarium oxysporum f. sp. cucumerinum J. H. Owen is one of the major destructive soilborne diseases and results in considerable yield losses. Methyl bromide was once the most effective disease control method but has been confirmed as harmful to the environment. Using suppressive media as biological controls to assist crop growth is becoming popular. In this study, Fusarium wilt of cucumber was successfully controlled by a newly identified suppressive media: vinegar residue compost-amended media (vinegar residue compost mixed with peat and vermiculite in a 6:3:1 ratio (v/v) vinegar residue substrate (VRS). Greenhouse experiments were carried out to evaluate the effect of VRS on the growth of cucumber seedlings and disease suppression. The control was peat/vermiculite (2:1, v/v). To identify the mixed media most suitable for the growth of plants and their suppressiveness indicators, we evaluated the biological characteristics of cucumber, the physicochemical and biochemical properties of the growth media, and the enzyme activities. Total organic C (C(org)), microbial biomass C (C(mic)), basal respiration (R(mic)), and enzyme (catalase, invertase, urease, proteinase, phosphatase, β-glucosidase, and hydrolysis of fluorescein diacetate) activities increased significantly after vinegar waste compost amendment. The compost media also showed a significantly positive effect on the growth of cucumber seedlings and the suppression of the disease severity index (DSI, 38% reduction). The cucumber rhizosphere population of F. oxysporum f. sp. cucumerinum (FOC) was significantly lower in VRS than in the control. These results demonstrate convincingly that vinegar residue compost-amended media has a beneficial effect on cucumber growth and could be applied as a method for biological control of cucumber Fusarium wilt.