Stability analysis of rabies virus in the environment.

Rabies, a fatal zoonosis, remains a burden in developing countries. Concerns about indirect environmental transmission and appropriate post‑exposure prophylaxis (PEP) are increasing, yet data on RABV stability under various conditions are limited. In this study, we systematically evaluated the stability of RABV (CVS‑11 strain) on six surface (glass, rubber, plastic, polypropylene surgical mask, fabric, and paper) at 25℃, and its stability in brain tissue, muscle, and simulated saliva under three simulated seasonal conditions: summer (35℃, 75% RH), spring/autumn (15℃, 43% RH), and winter (4℃, 33% RH). Viral titers were determined by fluorescent focus assay, complemented by direct fluorescent antibody staining and qPCR for nucleic acid detection. On surfaces, RABV titers declined from about 107 FFU/mL to 103 FFU/mL over 72 h, with inactivation occurring significantly faster on fabric than on other materials. Under simulated summer conditions, no infectious virus could be recovered from saliva after 12 h or from brain and muscle tissues after 24 h, but viral RNA remained detectable by qPCR. However, under spring/autumn and winter conditions, the virus still exhibited relatively high titers at 24 h, with enhanced stability at 4℃. These findings provide an evidence‑based framework for diagnostic laboratories to interpret nucleic acid tests in degraded specimens and to establish sample acceptance criteria. By clarifying the limited time window of environmental infectivity, they also help alleviate rabies‑related anxiety and support more nuanced post‑exposure prophylaxis decisions.