This study aims to investigate the effects of nanoparticles of zinc oxide (Nano-ZnO) supplementation on productive performance, nutrient digestibility, mineral retention, and blood biochemistry of Californian male rabbits under hot climatic conditions. A total of 100 Californian male rabbits 90 days of age (BW: 1790 ± 13 g) were randomly assigned to one of five treatment diets: Nano-ZnO at 0, 25, 50, 75, or 100 mg/kg, respectively, for 90 days. The rabbits were raised in an open house system (average ambient temperature of 39 °C, relative humidity of 30–35%, and temperature-humidity index of 33.6–34.0). Each treatment had 20 rabbits, and each rabbit was considered as a replicate. During the entire study period, there were no significant differences (P > 0.05) in BW, BW gain, feed intake, or feed conversion ratio between the treatment groups. Nano-ZnO supplementation at 25, 50, 75, and 100 mg/kg enhanced the digestibility coefficient of crude protein (CP) and ether extract (EE) considerably (quadratically, P < 0.05) when compared to the control group. In addition, when Nano-ZnO was supplemented at 25, 50, 75, and 100 mg/kg, serum levels of creatinine, alanine aminotransferase (ALT), and aspartate aminotransferase (AST) were considerably lower than the non-supplemented control group. Furthermore, when comparing the control and Nano-ZnO-fed rabbits, serum testosterone concentrations increased significantly (P < 0.05) in the treated groups. In addition, compared to the other groups, the 50 mg/kg Nano-ZnO group had the highest cecal Lactobacilli spp. count. These findings suggest that supplementing Californian male rabbits with 50 mg/kg dietary Nano-ZnO enhanced CP and EE digestibility, cecal Lactobacilli spp., as well as serum testosterone levels, and decreased serum ALT and AST levels under hot climatic conditions.