Usutu virus in blackbirds (Turdus merula) with clinical signs, a case study from northern Italy

Usutu virus (USUV) is a mosquito-borne virus belonging to the family Flaviviridae, genus Flavivirus. Natural transmission cycle of USUV involves mosquitoes and birds, so humans and other mammals are considered incidental hosts. In this study, USUV infection was diagnosed in all wild blackbirds, collected from July to September 2018 in a wildlife recovery center in the province of Bologna, in the Emilia-Romagna region, northern Italy. All blackbirds showed neurological clinical signs, such as overturning, pedaling, and incoordination. Moreover, the subjects died shortly after arriving at the hospitalization center. Virological investigations were performed by real-time PCR on frozen samples of the spleen, kidney, myocardium, and brain for the detection of Usutu (USUV) and West Nile (WNV) viruses. The small and large intestine were used as a matrix for the detection of Newcastle disease virus (NDV). All 56 subjects with neurological clinical signs were positive for USUV, only one subject (1.8%) tested positive for WNV, and no subject was positive for NDV. The most represented age class was class 1 J (58.9%), followed by class 3 (25.0%), and lastly from class 4 (16.1%). Most of the blackbirds before dying were in good (51.8%) and fair (39.3%) nutritional status, while only five subjects (8.9%) were cachectic. The USUV genomes detected in the blackbirds of this study fall within the sub-clade already called EU2 that has been detected since 2009 in the Emilia-Romagna region. Neurological clinical signs in USUV-affected blackbirds are still widely discussed and there are few works in the literature. Although our results require further studies, we believe them to be useful for understanding the clinical signs of Usutu virus in blackbirds, helping to increase the knowledge of this zoonotic agent in wild species and to understand its effect on the ecosystem. The goal of this study was to report—in the context of the regional passive surveillance program—the detection of USUV RNA in its most important amplifying host, the common blackbird, when showing clinical signs before death.