In the study, zinc oxide nanoparticles (ZNOPs) at concentrations of 20, 40, and 60 mg/kg were tested for their antimicrobial action against the oocysts of Eimeria tenella. The oocysts of E. tenella were isolated from the feces of broilers received at the veterinary hospital in Diwaniyah Province and initially diagnosed by compound optical microscopy. The oocysts were confirmed molecularly by polymerase chain reaction targeting the ITS1 gene with a molecular weight of 409 bp. The results in the first week showed that ZNOP concentrations of 20 and 40 mg/kg possess various activities against E. tenella, while 60 mg/kg was the most effective in reducing excreted oocysts compared to the positive control and amprolium group, along with the appearance of mild symptoms and a mortality rate of 0.8%. In the second week of infection, excreted oocysts and mortality rates generally decreased in all treated groups. A comparison of all groups showed that the 60 mg/kg ZNOP-treated group had a significantly lower number of excreted oocysts, and all birds in this group recovered during the second week of infection. These findings revealed the prospect of using ZNOPs against E. tenella in challenging situations of the appearance of resistance to anticoccidial agents.