Some plants have been interested as important sources in which the drugs including antimicrobial agents can be prepared. This study aimed to evaluate effects of the laser radiation on the Chloroform crude extract that was produced from Eucalyptus species leaves. Both extracts either laser-exposed crude chloroform extract and non-laser exposed one were in vitro tested against the growth of some pathogenic fungal species were Candida albicans, Candida parapsilosis, Aspergillus fumigatus, Microsporum canis, and Trichophyton canis  isolated from animal obtained from isolate bank of Veterinary Microbiology laboratory/ Collage of the Veterinary Medicine/University of Mosul. The results of this study showed the laser-exposed Chloroform crude extract produced inhibitory zones against C. parapsilosis and M. cains that the observed inhibitory zones were 15 and 10 mm, respectively but the non-laser exposed extract resulted in 10 mm against C. parapsilosis only. In this context, ketoconazole was also used as positive control and inhibited growth of the C. albicans and A. fumigatus only to give 20 and 18 mm against these fungi, respectively. Both extracts appeared compounds in their contents using GC-MS which showed three similar compounds in them were methanesulfonylacetic acid, pyrimidine-4,6(3H,5H)-dione, 2-butylthio- , and hexanedioic acid, bis(2-ethylhexyl) ester. Increasing antifungal activity of these compounds after exposure to laser radiation must be taken in attention. Although, there were no remarkable inhibitory results on the other fungi, the exact mechanism as antifungal is unclear and needs to be clarified in the future.