Evidence for West Nile Virus and Usutu Virus Infections in Wild and Resident Birds in Germany, 2017 and 2018.

Wild birds play an important role as reservoir hosts and vectors for zoonotic arboviruses and foster their spread. Usutu virus (USUV) has been circulating endemically in Germany since 2011, while West Nile virus (WNV) was first diagnosed in several bird species and horses in 2018. In 2017 and 2018, we screened 1709 live wild and zoo birds with real-time polymerase chain reaction and serological assays. Moreover, organ samples from bird carcasses submitted in 2017 were investigated. Overall, 57 blood samples of the live birds (2017 and 2018), and 100 organ samples of dead birds (2017) were positive for USUV-RNA, while no WNV-RNA-positive sample was found. Phylogenetic analysis revealed the first detection of USUV lineage Europe 2 in Germany and the spread of USUV lineages Europe 3 and Africa 3 towards Northern Germany. USUV antibody prevalence rates were high in Eastern Germany in both years. On the contrary, in Northern Germany, high seroprevalence rates were first detected in 2018, with the first emergence of USUV in this region. Interestingly, high WNV-specific neutralizing antibody titers were observed in resident and short-distance migratory birds in Eastern Germany in 2018, indicating the first signs of a local WNV circulation.

West Nile Virus and Usutu Virus Monitoring of Wild Birds in Germany.

By systematically setting up a unique nation-wide wild bird surveillance network, we monitored migratory and resident birds for zoonotic arthropod-borne virus infections, such as the flaviviruses West Nile virus (WNV) and Usutu virus (USUV). More than 1900 wild bird blood samples, from 20 orders and 136 different bird species, were collected between 2014 and 2016. Samples were investigated by WNV and USUV-specific real-time polymerase chain reactions as well as by differentiating virus neutralization tests. Dead bird surveillance data, obtained from organ investigations in 2016, were also included. WNV-specific RNA was not detected, whereas four wild bird blood samples tested positive for USUV-specific RNA. Additionally, 73 USUV-positive birds were detected in the 2016 dead bird surveillance. WNV neutralizing antibodies were predominantly found in long-distance, partial and short-distance migrants, while USUV neutralizing antibodies were mainly detected in resident wild bird species, preferentially with low seroprevalences. To date, WNV-specific RNA has neither been detected in wild birds, nor in mosquitoes, thus, we conclude that WNV is not yet present in Germany. Continued wild bird and mosquito monitoring studies are essential to detect the incursion of zoonotic viruses and to allow risk assessments for zoonotic pathogens.