Background Infectious laryngotracheitis (ILT) and infectious bronchitis (IB) are two common respiratory diseases of poultry that inflict great economic burden on the poultry industry. Developing an effective agent against both viruses is a crucial step to decrease the economic losses. Therefore, for the first time green synthesized silver nanoparticles using Cyperus rotundus L. aqueous extract was evaluated in vitro as a potential antiviral against both viruses. Results Silver nanoparticles from Cyperus rotundus were characterized by the spherical shape, 11–19 nm size, and zeta potential of − 6.04 mV. The maximum nontoxic concentration (MNTC) was 50 µg mL−1 for both viruses without harmful toxicity impact. The study suggested that some of the compounds in C. rotundus extract (gallic acid, chlorogenic acid, and naringenin) or its silver nanoparticles could interact with the external envelope proteins of both viruses, and inhibiting extracellular viruses. Conclusions The results highlight that C. rotundus green synthesized silver nanoparticles could have antiviral activity against infectious laryngotracheitis virus (ILTV) and infectious bronchitis virus (IBV) in chickens. Graphical Abstract