Leptospirosis caused by spirochete bacterium of genus Leptospira affects humans and animals worldwide. Rodents are major reservoirs of leptospires whereas wetland and aquatic migratory birds also carry and transmit leptospires. Leptospirosis studies in fish are lacking in African countries despite favourable environment and abundant reservoirs, which can spread leptospires into aquatic habitats and infect fish. The objectives of this study were to determine presence of Leptospira in fish; the prevalent Leptospira serovars and whether are related to serovars reported in animals; and to ascertain potential public health risk. Live tilapia, catfish and eel fish (n=48) were caught in Mindu Dam in Morogoro Municipality in eastern Tanzania. Blood sample was collected using syringes and needles to obtain serum for serological detection of leptospirosis using gold standard microagglutination test utilizing local and reference Leptospira serovars Sokoine, Kenya, Pomona and Hebdomadis. Twenty-six fish (54.2%) were positive for serovar Kenya (29.2%) and Sokoine (25%). Leptospira prevalence was high in both catfish (58.3%) and tilapia fish (47.8%). Thus, different fish types are infected with Leptospira found in animals. Fish could be source of Leptospira infection to humans since tilapia and catfish are the common fish type widely consumed in Tanzania. Further study covering lakes, rivers and dams is required to better understand the prevalence of Leptospira in fish and actual public health threats.