Buffaloes are one of the important farm animals in the south of Iraq and play an essential economical role mainly acting as dairy, meat, and draft animals. This study intended to diagnose buffalo mycotic eye infections in Thi-Qar province/Iraq. Some 250 buffaloes in the herd of 3,700 animals suffered from eye infections from December 2017 to November 2018. Eye swabs were collected from each infected eye of the affected buffaloes of both sexes before treatment. The animals were in different age groups. All samples were transferred to the laboratory in transfer media, and cultured on Sabouraud dextrose (SDA) agar with and without 0.05 g/mL and 0.4 g/mL chloramphenicol and cycloheximide, respectively. Later, the agars were incubated at 25o C and 37o C. The total percentage of eye infection was (6.75%), constituting (49.2%) mycotic infections. The predominant clinical manifestations that appeared on the infected buffaloes were eye inflammation represented by congestion, lacrimation, the opacity of cornea and edema, and reduced productivity of the infected animals. Different fungal isolates were identified from the samples including Aspergillus fumigates, Aspergillus flavus, Aspergillus niger, Penicillium spp., Alternaria spp., Fusarium spp., Candida spp., Cladosporium spp., Rhodotorula spp., Mucor spp. and Rhizopus spp. Calves buffaloes below one-year-old were more prone to mycotic infection than one-year-old or more. Additionally, male buffaloes were more susceptible to infection than females. In conclusion, this study isolated various types of fungus from the inflamed eyes of buffaloes. Fungal eye infection and the potential risk factors for fungal keratitis in buffaloes were also investigated. The study also approved the rapid diagnosis of fungi by direct microscopic detection and culture. The author recommends future studies including large numbers of the buffalo herd in Iraq to determine the epidemiology of this condition in the country.