AimsTo identify potential biological control agents against Verticillium wilt in olive through a mass screening approach. Method and ResultsA total of 47 strains and nine mixtures of micro-organisms were evaluated against Verticillium dahliae in a three stage screening: (i) invitro, by the effect on the mycelial growth and spore germination of the pathogen; (ii) in natural infested soil, by the effect on the reduction of microsclerotia of the pathogen; (iii) in planta, by the effect on the infection of olive plants under controlled conditions. Various fungal and bacterial strains and mixtures inhibited the pathogen and showed consistent biocontrol activity against Verticillium wilt of olive. ConclusionThe screening has resulted in promising fungi and bacteria strains with antagonistic activity against Verticillium, such as two non-pathogenic Fusarium oxysporum, one Phoma sp., one Pseudomonas fluorescens and two mixtures of micro-organisms that may possess multiple modes of action. Significance and Impact of the StudyThis study provides a practical basis for the potential use of selected strains as biocontrol agents for the protection of olive plants against V.dahliae infection. In addition, our study presented an effective method to evaluate antagonistic micro-organisms of V.dahliae in olive.