Using vegetable protein sources as a replacement for fish meal (FM) in the diet of Pacific white shrimp (PWS) has a negative impact on their health. Acute hepatopancreatic necrosis disease (AHPND), caused by Vibrio parahaemolyticus, affects PWS and causes financial losses. Nucleotides modulate the immune response and could contribute to counteracting these issues. Our objective was to evaluate the effects of nucleotide supplementation on performance, immune response, and survival when challenged with V. parahaemolyticus, in PWS receiving a diet with FM partially replaced with vegetable protein sources. A feeding trial (1000 PWS; 56 days) and a challenge trial (600 PWS; 10 days) were performed using diets with different FM inclusion levels (26%, 23.4%, 22.1%, and 20.8%), with or without 0.1% nucleotides. A non-challenged, non-supplemented group was also used in the challenge trial. Adding nucleotides to diets with reduced FM allowed significantly better results in growth performance parameters and total hemocyte count (THC). In the challenge trial, compared to control, nucleotide supplementation led to significantly higher THC and survival rate 15 h post-challenge. In conclusion, adding nucleotides to PWS diets improves their immune response and resistance to aquatic pathogens, allowing FM to be replaced by vegetable protein sources without negatively affecting performance.