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¹ Bursa Uludağ University, Faculty of Agriculture, Animal Science Department, 16059 Bursa, Türkiye DOI : 10.34233/jpr.1597124 - In this study it was aimed to investigate effects of ozone treatment during incubation of broiler eggs as an alternative application for fumigation. A total of 240 eggs from 45-week-old Ross 308 broiler breeder flocks were utilized. The eggs were weighed and randomly assigned to two groups: one without ozone and one with ozone. A commercial ozone generator was placed in the incubator, with ozone gas generated for 1 minute every hour to provide ozone gas at a concentration of 0.050 ppm. Throughout the 18-day incubation phase, ozone gas was applied in three-day cycles. On the 18th day of incubation, six randomly selected eggs from each experimental group were placed in sterile bags to assess the microbial load of the eggshell. Furthermore, six eggs were randomly selected from each experimental group to assess embryo growth. The total aerobic bacteria and Coliform sp. count were found higher in control than ozone group (P < 0.01). The yeast mold count, egg weight, embryo weight and embryo yolk sac weight were found to be similar between the groups (P > 0.05). It can be determined that using ozone instead of fumigation during incubation in broiler chicken eggs reduces eggshell total aerobic bacteria and coliform counts, but not affected to embryo traits such as weight, yolk sac weight and length. Keywords : Ozone Incubation Fumigation Eggshell microbial load Broiler