This study aimed to assess the effectiveness of using lavender essential oil to sanitize fertile eggs instead of formaldehyde; on the eggshell bacteria count and quality of chicks. Under aseptic circumstances, 825 white eggs (67.93±0.63 g) were collected from 60 weeks old (Lohmann White breeder chickens) and randomly divided into five treatments (non-sanitized and sanitized with distilled water, lavender essential oil, Glutaraldehyde GPC-8, and formaldehyde fumigation) before incubation and at 10th day of incubation. The results showed that according to the fertile eggs, non-sanitized and formaldehyde groups was reduced hatchability compared with other treatments. All sanitized groups reduced the dead embryo compared with non- sanitized. Total aerobic bacterial counts on the hatching eggshell surface were significantly reduced due to using Lavender essential oil and GPC8 compared with formaldehyde, non-sanitized, and sanitized with distilled water before setting in an incubator. Also, at the 18th day of hatching eggs, spraying with lavender essential oil reduced significantly the number of total aerobic bacteria compared to all other treatments. Lavender essential oil treatment recorded the highest chick quality compared with all other treatments except Glutaraldehyde, and the lowest chick quality recorded for the formaldehyde group. Also, the abnormal chicks were significantly reduced with Lavender essential oil compared with formaldehyde. Eggs for incubation can be safely and effectively treated with lavender essential oil. It is strongly advised to use it in place of formaldehyde and other toxic compounds while sanitizing fertile eggs.