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Continental waters salinisation is a global threat that has grown because of climate change and human activities, but little is known about how and what biological tracts are affected. The aim of this study was to investigate the influence of different water salinities on the expression of HSP70, PCNA and caspase-3 during spermatogenesis of Nile tilapia. Adult males were submitted to four salinity treatments: (S-0) fresh water, (S-7) 7 g L-1, (S-14) 14 g L-1, and (S-21) 21 g L-1 for 1, 4, and 9 days. All specimens were in spermatogenic activity and the highest values of the gonadosomatic index (GSI) occurred in the S-0 and S-7. In the morphometric analysis, spermatocytes were the most frequent germ cell detected in all treatments (>50%) and spermatids achieved about 20% of the testicular proportion, with few variations among treatments. Spermatozoa were significantly reduced only in S14 compared to S7. Leydig cells were significantly increased in S-14 when compared to S7 but plasma concentrations of 11-KT showed no significant difference among treatments. ELISA assay showed higher testicular expression of HSP70 at 1 day in all groups, followed by a significant decrease at days 4 and 9 in S-14 and S-21. The expression of PCNA was significantly lower while the activity of caspase-3 was higher in S-14 and S-21 when compared to So and S7. These results indicate that higher salinities in S-14 and S-21 interfere with the relationship between testicular HSP70, PCNA, and caspase-3, but with few effects over spermatogenesis dynamics of Nile tilapia. Published by Elsevier Inc.