A microbiological study of organisms associated with spoilage of fresh fish samples collected from two ponds located in Benin City, Nigeria was carried out. Samples from the fish skin, gills and flesh were cultured in three media; nutrient agar, Maconkay agar and potato dextrose agar and on examination the presence of five bacteria species namely; Staphylococcus aureus, Klebsiella sp., Salmonella sp., Escherichia coli, Pseudomonas sp., and four fungi species namely; Aspergillis niger, Geotrichum sp., Rhizopus sp. and Pennicillium sp. was confirmed. The highest colony count was obtained from the skin samples in all the media. Antibiotic sensitivity pattern showed that all isolates were resistant to Gentamicin and Amoxicillin and the presence of the above pathogens in the fresh fish samples could pose a potential public health threat especially to consumers. It is recommended that better handling and processing methods should be adopted to reduce or eliminate health risk to fresh fish consumers.