Background/Aim: Some of the antifungal drugs used in the current treatments regime are responding to antimicrobial resistance. In rural areas of Southern Tanzania, indigenous people use antifungal drugs alone or together with medicinal plants to curb the effects of antibiotic resistance. This study documented ethnobotanical information of medicinal plants used for managing fungal infections in the Southern Highlands of Tanzania and further assess their safety. Materials and Methods: Ethnobotanical survey was conducted in Makete and Mufindi districts between July 2014 and December 2015 using semi-structured questionnaires followed by two focus group discussions to verify respondents' information. Cytotoxicity study was conducted on extracts of collected plants using brine shrimp lethality test and analyzed by MS Excel 2013 program. Results: During this survey about 46 plant species belonging to 28 families of angiosperms were reported to be traditionally useful in managing fungal and other health conditions. Among these, Terminalia sericea, Aloe nutii, Aloe lateritia, Zanthoxylum chalybeum, Zanthoxylum deremense, and Kigelia africana were frequently mentioned to be used for managing fungal infections. The preparation of these herbals was mostly by boiling plant parts especially the leaves and roots. Cytotoxicity study revealed that most of the plants tested were nontoxic with LC50 > 100 which implies that most compounds from these plants are safe for therapeutic use. The dichloromethane extract of Croton macrostachyus recorded the highest with LC50 value 12.94 mu g/ml. The ethnobotanical survey correlated well with documented literature from elsewhere about the bioactivity of most plants. Conclusions: The ethnobotanical survey has revealed that traditional healers are rich of knowledge to build on for therapeutic studies. Most of the plants are safe for use; and thus can be considered for further studies on drug discovery.