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Sachet water is the ultimate source of drinking water in Nigeria. Such waters are required to comply with minimum standards in order to protect public health as safe drinking water. A study was conducted to investigate the presence and effects of fungi in sachet water in University of Benin Teaching Hospital using five brands of sachet water samples. The sachet waters were subjected to pour plate method to determine the fungal content of the water. All samples (OWC, Superfine, Notre Dame, Rancho and Olivia sachet water) tested positive for the presence of fungi. OWC sachet water was the most infected with 57% of fungi isolate, while superfine was the least infected with 28.5% of fungi isolate. The fungal isolates were identified as Aspergillus niger, A. flavus, A. tubigensis, Botrytis cinerea, Pythium insidium, Mucor sp and Rhizopus sp. Aspergillus sp and Mucor sp were the most prevalent as they were present in all the sachet water samples while Botrytis cinerea and Pythium insidium were the least prevalent as they present only in OWC sachet water. These fungi have the potential to cause allergic reactions or diseases in humans.