ABSTRACT The present study aimed to identify yeasts species isolated from fish feed and the intestinal tracts of tambatinga fish (Colossoma macropomum × Piaractus brachypomum) cultivated in a Brazilian fish farm. Twenty tambatingas and 30 fish feed samples from different brands were acquired from two commercial establishments in the state of Piauí, Brazil. The sampled guts were divided into three equal parts, namely the anterior, medium and posterior portions, totaling 60 samples. Molecular identification was performed by PCR amplification and sequencing of the D1/D2 regions of the large rRNA subunit gene. Sixteen Candida nivariensis strains in the gut were identified. The fish feed samples showed a higher variability of yeast species, with the following species isolated: Hyphopichia burtonii (23.08%), Lodderomyces elongisporus (15.39%) and Trichosporon asahii (15.39%), Candida nivariensis (7.69%), C. orthopsilosis (7.69%), C. parapsilosis (7.69%), Rhodotorula minuta (7.69%), Sterigmatomyces elviae (7.69%), Cryptococcus liquefaciens (7.69%). Yeast found in feed, however, was not always isolated from tambatinga gut. Molecular identification allowed for the isolation of yeast species not previously reported in fish feed and gut samples.