The ten most frequently isolated fungi from the wood of the dead branches of Acer pseudoplatanus L. were tested in dual cultures to evaluate their in vitro antagonistic activity against Eutypella parasitica R.W. Davidson and R.C. Lorenz, the causative agent of a destructive disease of maples in Europe and North America. The tested fungi, treated also as challenge isolates, were Diaporthe sp., Eutypa sp., Eu. maura, E. parasitica, Fusarium avenaceum, Neocucurbitaria acerina, Neonectria sp., Peniophora incarnata, Petrakia irregularis, and Phomopsis pustulata. The antagonistic ability of each challenge isolate was evaluated by calculating an index of antagonism (AI) based on the interaction type in the dual cultures. The results of competition between the fungal isolates were quantified after re-isolations from the interaction zone (s). The dual cultures revealed two main types of competitive interactions: Deadlock, consisting of mutual inhibition after mycelial contact or at a distance, and replacement, reflecting in the inhibition of E. parasitica, followed by partial overgrowth by the replacing fungus. Statistical analysis showed significant differences in average AI and s of challenge isolates between different dual culture assays. Based on the results of the antagonism index, Eutypa sp., Eu. maura, Neonectria sp., and P. incarnata had the highest inhibitory effect on E. parasitica growth and were recognized as the most promising candidates for further biocontrol studies of E. parasitica. The mycelium of E. parasitica at the interaction zones remained mostly viable, except in dual cultures with Eutypa sp., F. avenaceum, and Neonectria sp., where re-isolations did not yield any colony of the E. parasitica isolate. Based on the results, we assume that E. parasitica is a weak competitor, which invests less energy in direct mycelial competition. We discuss the potential of the observed antagonists as a possible biocontrol of Eutypella canker of maple. Nevertheless, additional experiments should be performed for a solid conclusion about competitive ability of E. parasitica and usefulness of antagonists as biocontrol.