The effect of temperature (18˚C -30˚C), water activity (0.85 -1) and pH (4 -9) was studied by dual culture technique on the antagonism of Bacillus amyloliquefaciens and Trichoderma harzianum to Colletotrichum acutatum, responsible of strawberry (Fragaria x ananassa (Weston) Duchesne ex Rozier) anthracnose. The antagonistic bacteria's strains behave significantly and differently according to the parameters studied. These results reveal useful information about the applicability of their biocontrol in agricultural culture with the change of environmental factors.