A medical establishment such as hospitals and medical centers generate a sizable amount of hazardous waste. In Yemen, the hospitals' wastes are still largely mismanaged, mainly because the sector did not know what to do with the waste. The present study was undertaken to determine the bacterial and fungal agents present in different various of the hospitals' solid waste. The samples were collected from the different wards/departments and from the central storage rooms/dustbins of the hospitals in Sana'a city, Yemen. The presence of bacterial and fungal agents in the hospitals' waste was determined using the conventional bacteria, and fungi identification methods. The bacteria and fungi were detected in the hospitals' solid waste were Klebsiella spp. (9.3%), E. coli (12.7%), Citrobacter spp. (8.5), Candida spp. (18.6%), Proteus spp. (9.3%), Cladosporium werneckii spp. (19.5%), Bacillus spp. (9.3%), Aspergillus spp. (7.6%), Trichothecium spp. (0.8%), Mucor spp. (3.4%), and Acinetobacter spp. (0.8%). This study results confirmed that the prevalence types of microorganism vary based on seasonal and spatial variables