In Colombia, economic losses due to attack of Fusarium oxysporum in the gooseberry plantation are considerable. Fungi and bacteria isolated from 2 hen manure sources were evaluated for their potential as biological control agents of this pathogen. The evaluation was conducted in Petri dishes containing PDA by placing a 5 mm diameter disk, colonized by this pathogen, in the center of the plates and at 3 cm from the center, over the horizontal and vertical axis, each of the microorganisms isolated from the hen manure. The isolated microorganisms that showed antagonism were later evaluated in vitro for their capacity to inhibit growth and sporulation of F. oxysporum. Each of the microorganisms that showed the greatest antagonism potential was simultaneously inoculated with the pathogen in gooseberry seedlings and evaluated for their effects against the incidence of the disease and the death of the seedlings. The results indicated that of the 39 microorganisms isolated from the pure hen manure, 6 showed antagonism against F. oxysporum and among them the most effective to inhibit in vitro its growth and sporulation were the fungi H2 and H6 and the bacteria B17 and B19. The bacteria B17 and B19 resulted to be the most effective in reducing not only the incidence, but also the death of the seedlings caused by the pathogen. According to the identification results, the H2 and H6 fungi belong to the genera Geotrichum and Trichoderma, respectively and the B17 and B19 bacteria to the genus Bacillus.