Zinc oxide nanoparticles (ZnO NPs) are widely used in diverse fields. However, the toxicological effects of ZnO NPs remain inadequately explored. This study aimed to investigate the toxicities of various concentrations of ZnO NPs <50 nm (ZnO NP50), <100 nm (ZnO NP100), and ZnO bulk particles (ZnO BP) after different exposure durations using Caenorhabditis elegans (C. elegans) as an in vivo model, focusing on reproductive capacity, feeding behavior, and lifespan. Polymerase chain reaction (PCR) was conducted to assess expressions of stress-related cep-1 and pmk-1 genes. The results revealed that the effects of ZnO particles on C. elegans’ reproductive capacity were inconsistent after exposure for 24, 48, and 72 h. It was seen a general concentration-dependent reduction in pharyngeal pumping rate, with 1000 [Formula: see text]g/mL having the most significant impact across all ZnO particle treatments. Lifespan studies showed no significant differences post exposure of ZnO particles for the mean lifespan of C. elegans, while significant differences for ZnO NP50 treatment at 100 [Formula: see text]g/mL and 1000 [Formula: see text]g/mL compared to the control and the ZnO BP groups were observed. Finally, PCR results suggested significant up-regulation of cep-1 and pmk-1 gene expression, showing a size-dependent linear trend with ZnO NP50 > ZnO NP100 > ZnO BP. In conclusion, this study underscores the complex size-dependent toxicological effects of ZnO NPs on C. elegans, highlighting significant impacts on pharyngeal pumping rate and gene expression profiles associated with stress response and DNA damage repair pathways. These findings contribute to our understanding of NP toxicity and underscore the importance of size, concentration, and exposure duration considerations in assessing their biological effects.