This study aims to characterize the spatial and seasonal distribution of the fish assemblage and evaluate the integrity of streams in a sustainable use area that includes integral protection conservation units in Distrito Federal, Central Brazil (Cerrado biome). For the study, 12 stretches of 8 streams were sampled in 2008 (dry season) and 2009 (wet season). For that evaluation was estimated the Physical Habitat Index (PHI), vegetation cover (VC), pH, dissolved oxygen, turbidity, and conductivity. We recorded 22 species, about eight undescribed species, by a total of 2,327 individuals. The most representative families in number of species were Characidae (31.8%), Loricariidae (31.8%), and Crenuchidae (13.6%). Knodus moenkhausii was the most abundant species with 1,476 individuals, added to Astyanax sp., Phalloceros harpagos, and Hasemania sp. they represent over 95% of the total abundance. The species Astyanax sp. (occurring in 79.2% of the stretches) and K. moenkhausii (50.0%) were considered constant in both seasons. The longitudinal gradient (River Continuum) exerts a strong influence on the studied assemblage. According to CCA, the variables that structure the fish assemblage are based on aspects related to water volume and habitat complexity. No seasonal variation in richness, diversity, abundance, and mass were detected. A cluster analysis suggests a separation of species composition between the stretches of higher and lower orders, which was not observed for seasonality. The streams were considered well preserved (mean PHI 82.9±7.5%), but in some stretches were observed anthropogenic influence, detected in the water quality and, mainly, on the riparian vegetation integrity. The exotic species Poecilia reticulata was sampled in the two stretches considered most affected by anthropogenic activities by PHI, conductivity, and VC.Esse estudo teve como objetivo caracterizar a distribuição espacial e sazonal da taxocenose de peixes e avaliar a integridade dos córregos de uma área de uso sustentável que inclui unidades de conservação de proteção integral no Distrito Federal, Brasil Central (bioma Cerrado). Para o estudo, foram amostrados 12 trechos de oito cursos d'água em 2008 (estação seca) e 2009 (estação chuvosa). Para essa avaliação foram utilizados o Índice Físico do Habitat (IFH), cobertura vegetal (CV), pH, oxigênio dissolvido, turbidez e condutividade. Foram coletadas 22 espécies, sendo oito não descritas, em um total de 2.327 indivíduos. As famílias mais representativas em número de espécies foram Characidae (31,8%), Loricariidae (31,8%) e Crenuchidae (13,6%). Knodus moenkhausii foi a espécie mais abundante, com 1.476 indivíduos, que juntamente com Astyanax sp., Phalloceros harpagos e Hasemania sp. representaram mais de 95% da abundância total. As espécies Astyanax sp. (ocorrendo em 79,2% dos trechos) e K. moenkhausii (50,0%) foram consideradas constantes nas duas estações. O gradiente longitudinal (rio Contínuo) exerce forte influência na taxocenose estudada. De acordo com a CCA, as variáveis que estru...