Objective: This work aimed to investigate the antimicrobial effect of traditional Saudi Arabian incense (white benzoin resin, Jawi) on indoor environment microbes in Shaqra University, Dawadmi Female Student Campus, and bacterial isolates from patients in Dawadmi General Hospital. Methods: The effect of white benzoin resin fumes was tested directly on clinical bacterial isolates. The bacterial culture plates were incubated with benzoin resin fumes within a confined space. The microbial quality of indoor air in four laboratories, lecture rooms, library, and restroom of the students was measured, and the effect of benzoin resin fumes on these indoor microbes was determined. The settle plate method with open Petri dishes containing different culture media was used to collect samples two times daily before and after benzoin resin fumes were used. Results: The growth of Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans was completely inhibited after exposure to benzoin resin fumes for 90 min. The growth of Klebsiella pneumoniae, Escherichia coli, and Enterobacter aerogenes was inhibited by 79%, 68%, and 55%, respectively, after the same incubation time. Fumigation of selected university premises with the incense decreased the total bacterial count from 23.5 Colony-Forming Units (CFU) to 6.25 CFU. Conclusions: The application of benzoin resin fumes in some university premises showed a promising effect on decreasing the total number of microorganisms in the air of some rooms and inhibiting the growth of some pathogenic bacteria.