The matrix effect of the thermophilic anaerobic digestion (TAD) process on inactivation of infectious laryngotracheitis virus (ILTV) was evaluated. Viral cell culture and real-time PCR were used for assessing removal of the viral infectivity and degradation of viral DNA, respectively. Results showed that the TAD-derived matrix alone can inactivate the virus and destroy the nucleic acid helix core of ILTV in a time-and- dose-dependent manner. No cytopathogenic effect (CPE) was observed in the cells exposed to ILTV pre-treated with TAD matrix for 1.5 h in experiment 1 and for 16 h in experiment 2. There was a significant statistical difference between TAD matrix treated and non-treated cultures (p < 0.001, Chi-test). Amplifiable ILT viral DNA was reduced 2.27 log by 1.5 h-treatment and was not present by 16 h-treatment with TAD matrix, indicating complete viral DNA fragmentation. The TAD process is an environmentally friendly way for disposing of poultry biowaste and carcasses. (C) 2012 Elsevier Ltd. All rights reserved.