Fungal toxins in feed are leading issue in poultry industry causing a detrimental effect on the performance and health of poultry. The study was carried out to determine the incidence and concentration of the aflatoxins and their major producer Aspergillus flavus in home mix layer poultry feed in respect of seasonal variation throughout the year. A total of (n?= 204) home mix poultry layer feed samples were analyzed for the isolation of fungi. The isolates were initially screened through colony morphology and microscopic examination. However, aflatoxin concentration was determined by ELISA. Revealed results indicated that, the highest percentage of A. flavus was found during the months of June to August 50/54 (92.5%) followed by September to November 43/65 (66.1%), March to May 21/40 (52.5%), and December to February 18/45 (40%). As a whole, the incidence was recorded 132/204 (64.7%). Moreover, of the 132 samples, 41 (31%) were exceeded in respect of aflatoxin contamination from the legal limit (20 μg/kg) imposed by Food Drug Association (FDA). Statistically, the growth of A. flavus and aflatoxin production was found significantly different in respect of seasonal variation. As highest total viable fungal count (9.9?×?10⁴ CFU/g) and aflatoxin level (72.27 μg/kg) were recorded during the months of June to August and lowest in December to February. Consequently, instantaneous essential control measures are demanded regarding appropriate storage and adequate drying in post-harvesting season. Along with surveillance plans and austere regulations for monitoring the aflatoxin contents for the wellbeing of consumers.