Developing novel formulations and processes to improve the fungal growth inhibition and biodegradability of NR products has become a major challenge for the scientific community, given the health risks associated with fungi in natural rubber (NR) products and the environmental concerns regarding NR waste. Consequently, this study comprehensively investigated the synergistic effects of chitosan and gamma irradiation on enhancing fungal growth inhibition and biodegradability in natural rubber latex (NRL) films. The research involved incorporating varying chitosan contents (0, 3, 6, or 9 phr) into NRL composites and exposing them to different gamma doses (0, 5, 10, or 15 kGy). The results showed that increasing the chitosan content and the gamma dose improved the ability of the NRL samples to inhibit fungal growth on their surfaces. This was evidenced by the absence of fungal colonies after 7 days of incubation on potato dextrose agar (PDA) plates for NRL samples irradiated at 5–15 kGy with 9 phr of chitosan. This determination was based on isolating fungi from the film surfaces, followed by serial dilution and a viable plate count. The biodegradability tests also revealed that the NRL films irradiated at 15 kGy with 9 phr of chitosan had the highest weight loss, reaching as high as 10.42 ± 0.62 % after soil burial for 8 weeks. However, the results indicated that gamma irradiation on the pristine NRL and chitosan/NRL films did not substantially alter the thermal stabilities, density, morphology, and functional groups of the samples. Lastly, by comparing the tensile properties of all the NRL films to the ASTM D3578-01a standard for examination gloves, the optimum conditions for the NRL films were 5 kGy of gamma irradiation with 9 phr of chitosan. This combination resulted in gloves with sufficient tensile strength and complete fungal growth inhibition.