Growth, survival, and use of feed of the F1 generation of two new strains of tilapia Oreochromis niloticus (GenBank Strain Y = OK078792 to OK078796 and Strain A = OK078797 to OK078807) produced by mass selection was compared with a known fast-growing commercial strain in Mexico (GenBank Strain Q = OR744820 to OR744838), during a grow-out phase of 165 days. Genetic variability including observed (Ho) and expected (He) heterozygosity, inbreeding coefficient (Fis), and paired comparisons of strain fixation index (Fst) of the three strains were also compared, and an additional analysis of molecular variance (AMOVA) was performed. Twelve hapas of 8.2 m3 (four for each strain) were stocked at a density of 25 org m−3 of 4.58 ± 0.67 g, which were fed with 45 to 25% crude protein feed under a dynamic feeding scheme of 8 to 3% live weight. Growth was estimated by growth rates in weight and length, and by the weight-specific growth rate (SGR). The use of feed was estimated by the total feed consumption rate and the feed conversion factor (FCF). Results indicate no significant differences (P > 0.05) in growth parameters of the three strains, except in SGR which was significantly higher in the control strain (Q) (2.75% day−1) as compared with strains Y and A (2.51 and 2.59% day−1 respectively). The average final weight and size of the fish were 314.6 ± 11.8 g and 25.5 ± 0.4 cm respectively. No significant differences (P > 0.05) were found in survival and feed utilisation among strains, with average values of 96.4% survival, feed consumption of 3.91 g day−1, and FCF of 2.08. Results of genetic diversity indicate that there were no significant differences (P > 0.05) in Ho and He of the three strains, with average values of 0.310 and 0.280 respectively. Fis values were negative and similar for the three strains, with a mean value of −0.109 which indicates no consanguinity in their populations. Finally, the paired comparisons of Fst were significantly different (P < 0.001), showing relatively low values but different from 0 (Y vs. A = 0.025, A vs. Q = 0.041 and Y vs. Q = 0.055), indicating slight but significant population differentiation between strains. In conclusion, the performance and genetic variability of the F1 generation of the two new strains suggest that valuable broodstock is now available for mass production of fry, with similar performance as to one of the best strains of tilapia in Mexico.