Abstract: Background and Objective: The possible health hazard associated with the traditional production of non-alcoholic beverages calls for regular microbiological quality control check to reduce the potential menace on humans’ health. The current study was planned to investigate the microbiological quality and antibiotic susceptibility pattern of bacteria associated with burukutu beverage sold in Akure metropolis with a view to establish its biosafety. Materials and Methods: This research was carried out between April and July, 2016. Total aerobic microbial counts were determined using pour plate technique on nutrient agar, MacCankey agar, potato dextrose agar and sabroud dextrose agar. Afterward, the counts were reported as colony forming unit per milliliter ( CFU mL–1) for bacteria and spore forming unit per milliliter (SFU mL–1) for fungi. The antibiotic sensitivity test was carried out using agar diffusion method. The susceptibility of the bacterial isolates to commercial antibiotics were determined by measuring the zone of inhibition in millimeter and interpreted according to Clinical and Laboratory Standards Institute (CLSI). Results: The highest bacterial and fungal loads of 35.0×109 and 105.0×109 SFU mL–1, respectively was obtained from the samples collected from the Army Barrack (Seller 3), while the samples collected from Road block (Seller 1) had the least values. The identified microbial isolates were: Bacillus subtilis, B. cereus, Staphylococcus aureus, E. coli, Lactobacillus fermentum, Streptococcus lactis, Aspergillus niger, Penicillum italicum, Rhizopus japonicum, Fusarium spp. and Saccharomyces cereviseae. Streptococcus lactis and Lactobacillus fermentum had the highest percentage occurrence of 60% from samples collected from Road block and Army barrack respectively, while Aspergillus niger had the highest percentage occurrence of 62.5% from the sample collected from Road block. Appreciable numbers of the bacterial isolates were sensitive to commercial antibiotics. The isolates Streptococcus lactis showed resistant to all the antibiotics except Amoxycillin and Androcephin. Escherichia coli, Bacillus subtilis, Streptococcus and Staphylococcus aureus showed resistant to Amoxycillin, Zinacef, Septrin and Streptomycin. Conclusion: The presence of antibiotic resistant bacteria and mycotoxin-producing fungi in the samples are of great health concern both to human and animals.