Aim: This study was conducted to comparatively investigate the soil and air microflora of selected locations within the Federal University of Akure (FUTA), Ondo State, Nigeria. Study design: Experimental design Place and Duration of Study: The study was conducted at The Federal University Technology, Akure (FUTA), Ondo State, Nigeria comprising; School of Sciences, School of Agricultural and Agricultural Technology, School of Engineering and Engineering Technology, School of Health and Health Technology, Microbiology department, FUTA Health Centre, postgraduate hostel, female hostels (main Jibowu and Jibowu annex 1) and the male hostels (Akindeko and Abiola hall of residence) between June and July, 2019. Methodology: Isolation of bacteria and fungi from soil samples and air was conducted using specified techniques. Antibiotic susceptibility test was conducted via agar disc diffusion technique. Plasmid analysis and curing was conducted via standard protocols. Results: Pseudomonas aeruginosa, Klebsiella pneumoniae, Escherichia coli, Proteus mirabilis, Bacillus cereus, B. subtilis, Staphylococcus aureus, S. saprophyticus were isolated as soil and air microflora while Candida albicans, Aspergillus fumigatus, with Fusarium oxysporium were isolated from soil as A. flavus, A. niger, Penicillium chrysogenum was implicated in air. Bacillus cereus and B. subtilis showed utmost resistance to ceftazidime, cefuroxime, cloxacillin, augmentin, ceftriaxone, whereas Enterobacter aerogenes and Proteus vulgaris were susceptible to gentamycin at 17.66±1.52 mm and 16.00±2.00 mm respectively. Staphylococcus aureus was sensitive to gentamycin and ofloxacin at 17.30±0.57 mm and 18.66±0.57 mm. The multidrug resistant (MDR) bacterial strains were positive for plasmid DNA with 10 kilobase pairs, but were sensitive to all the antibiotics after curing indicating plasmid-borne resistance. Conclusion: Findings from this study inferred possible health consequences of MDR soil and air microflora as potential threat to human health which necessitates proper sanitary practices across different sampling areas adopted for this study to reduce potential incidence of bacterial and mycotic infections.