Fufu (Cassava dough-like food) is one of Nigeria's most consumed staple foods, particularly in the southern parts of the country. This study aims to detect and quantify the microorganisms in cooked fufu sold in major Owo, Ondo State marketplaces. Freshly cooked fufu samples of about 50/gram wrapped in nylon were aseptically collected from sellers in three major Owo markets and transported to the Federal University of Technology, Akure (FUTA) Crop, Soil, and Pest Management Department lab for analysis. For bacteria, Nutrient Agar (NA) plates were incubated at 37+2ºC for 24 hours, whereas PDA plates for yeast and mould were incubated at 27+2ºC for 48 hours. Count and record the colonies generating spores (cfu/g) from viable bacterium cells and fungal spores per gram of each fufu sample after incubation. Purified cultures of each isolate were obtained through subcultures and stored at 4 ºC for biochemical testing. Cooked fufu samples found mostly lactic acid bacteria like Lactobacillus plantarum, L. fermentum, L. brevis, Leuconostoc mesenteroides, Streptococcus thermophiles, S. mutans, Pediococcus cerevisiae, Staphylococcus hominis, Bacillus pumilus, acidophilus, L. lactis, Micrococcus spp., B. subtilis, Diplococcus spp, S. epidermidis, and Bifidobacterium and two yeasts, Saccharomyces cerevisiae and Zygosaccharomyces spp. Some main markets in Owo metropolis sold cooked fufu samples with bacterial loads ranging from 0.2 to 2.3 × 107 CFU/g, and yeast loads from 1.6 to 4.55 × 107 CFU/g. Good hygiene can increase the safety and quality of cooked fufu sold at Owo main markets and lower the danger of food-borne illnesses.