The current study aimed to determine the effect of silver and chitosan nanoparticles of size 10 to 30 nm on the dead of lice in vitro and in vivo to determine the optimal time and concentration to combat chicken lice. 100 local chickens Gallus gallus domesticus were collected from Al-Diwaniyah province and 6 species of local chicken lice were isolated: Menacanthus stramineus, Menacanthus pallidullus, Menacanthus cornutus, Goniodes gigas, Cuclotogaster heterographus and Bonomiella columbae. The results of treating lice with chitosan and silver nanoparticles at concentrations (40, 60, and 80) mg/mL in vitro and at different periods (5, 10, 15, and 30) minutes after treatment showed that chitosan and silver nanoparticles at a concentration of 80 mg/mL are the most effective in killing lice. The dead rate of lice reached 100% after 15 minutes of treatment with chitosan nanoparticles and 100% in the case of silver nanoparticles after 30 minutes. The results of spraying chitosan and silver nanoparticles on the body of chickens infected with lice experimentally, based on the relative therapeutic efficacy within 30 minutes, indicated that silver nanoparticles were the most effective in completely killing lice in the group treated with a concentration of 80 mg/kg after 30 minutes, where the percentage of therapeutic efficacy was 96.7%. This was followed by chitosan nanoparticles at a concentration of 80 mg/kg, and the percentage of therapeutic efficiency was 91.5%. Chitosan and silver nanocomposite have a promising effect in the elimination of lice infestation in chickens.