Abstract
Paslahepevirus balayani (Hepatitis E virus, HEV) is an emerging zoonotic pathogen with a wide host range, yet its circulation in African equids remains poorly understood. This study provides the first serological investigation of HEV in donkeys in Algeria, aiming to assess exposure levels and identify potential infection-related risk factors. Between 2019 and 2024, 183 donkeys were sampled across three northeastern provinces (Mila, Souk-Ahras, and Tébessa), representing diverse agroecological conditions and including both working donkeys and animals used in cross-border smuggling. Individual data on age, sex, health status, body condition, movement history, and season were recorded. Sera were screened using a proprietary recombinant antigen, which is highly conserved across different HEV strains. All samples tested negative for anti-HEV antibodies. The sample size exceeded the minimum required to detect the highest seroprevalence previously reported in donkeys (12.22%), supporting the epidemiological significance of the negative findings. The absence of seropositivity may reflect limited regional exposure to known HEV reservoirs, local husbandry practices, and environmental conditions that reduce infection risk. These results suggest that HEV is currently not circulating at detectable levels in donkeys from northeastern Algeria. Broader studies, including molecular approaches and additional species, are recommended to clarify HEV ecology in the region.