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Recently, donkeys have gained popularity mainly due to the use of donkey milk by the cosmetic industry and for human consumption. Gastrointestinal strongyles (GIS) are considered a potential cause of disease and reduced production in infected donkeys. European laws limit the use of anthelmintic drugs for the control of GIS in dairy donkey farms, thus the need to develop alternative control methods. This study aimed to test the in vitro inhibiting effects of three chitin degrading fungi (<i>Scopulariopsis brevicaulis</i>, <i>Metarhizium anisopliae</i>, and <i>Beauveria bassiana</i>) on the hatch and viability of donkey GIS eggs by using the egg hatch test, and to compare their activity to that of <i>Pochonia chlamydosporia</i>. About 150 eggs were added to 0.5 mL of sterile saline solution containing about 1.4 × 10⁸ spores of each fungal species or with 0.5 mL of sterile saline solution only (untreated controls). After incubation, the percentage of egg hatch reduction was calculated, and data were statistically analyzed. All fungi were able to significantly reduce (<i>p</i> < 0.05) the hatch of GIS eggs compared to the untreated controls. Further studies that aim to investigate the efficiency of these fungi in reducing donkey GIS eggs in contaminated environments are encouraged.