The poultry sector in Ghana has the potential to contribute significantly to domestic food and nutrition security. There is limited information regarding the knowledge of feed industry stakeholders on the presence of mycotoxins in poultry feed. This work was aimed at assessing the prevalence of aflatoxin and fumonisin in poultry feed, and mycotoxins awareness among stakeholders of the poultry feed value chain in Ghana. Three hundred and fifty (350) respondents consisting of 255 commercial poultry farmers, 85 feed processors and 16 feed vendors were involved in the study. AgraStrip® Quantitative test from RomerLab, USA was used to quantify total fumonisin and aflatoxin levels in the prepared feed samples. The study revealed that commercial feed producers, feed mills operators and on-farm feed processors are the three main sources of feed and ingredients for commercial poultry farmers in Ghana. The study identified a high level of mycotoxin contamination of feed and ingredients. Feed samples from the commercial feed processors recorded significantly lower aflatoxin levels (21.63+7.05 ppb, p<0.008). However, the threshold of 15 ppb for the Ghana Standards Authority, FAO/WHO maximum permissible limit of 30 ppb and EU regulatory limit of 20 ppb for poultry feeds were exceeded by 70% of all the samples. Fumonisin levels were less than the 30mg/kg FDA guidance levels for maize and maize by-products intended for consumption by breeding poultry. From the study, a majority of the farmers (85%), vendors (87.5%) and all of the feed processors have their own feed storage facilities or warehouses. Storage problems identified by these stakeholders included pest infestation and mold development, which usually occur after a few weeks of storage to the end of the storage period. Sanitation was poor along the feed chain, particularly during feed formulation and storage. Awareness of mycotoxins and their effect on animal and human health was low among the respondents. Some of those who have heard about aflatoxins were not aware of what it is and its effect on the health of humans and animals. This threatens the management of mycotoxins in the poultry feed chain