Evaluation of henipavirus chemical inactivation methods for the safe removal of samples from the high-containment PC4 laboratory

•Fixation methods for safe removal of samples from high containment laboratories.•10 % neutral buffered formalin inactivates Nipah virus within tissues ≤48 h.•4 % paraformaldehyde inactivates henipavirus-infected cell monolayers ≤15 min. Henipaviruses, Hendra (HeV) and Nipah (NiV), are highly pathogenic zoonotic agents that pose a serious health risk to human life, and as such are restricted to physical containment 4 (PC4) laboratories. For further analysis of virus-infected biological specimens, it is necessary to ensure absolute inactivation of any infectious virus present before removal from the PC4 laboratory. To evaluate the inactivation of HeV and NiV within infected samples, two chemical inactivation methods were assessed. Henipavirus-infected cell monolayers treated with 4 % paraformaldehyde (PFA) showed the complete inactivation of infectious virus, with an inactivation period of 15 min resulting in more than 8-log decrease in infectious titre. NiV-infected tissue samples treated with 10 % neutral-buffered formalin (NBF) showed a complete reduction of infectious virus in 7/8 ferret organs incubated for 24 h, with the remaining tissue demonstrating complete virus inactivation after 48 h. The chemical inactivation methods described herein evaluated two simple methods of henipavirus inactivation, resulting in the complete inactivation of infectious virus – an essential requirement for the safe removal and handling of biological samples from the PC4 laboratory.