**4. Results**

A fixed number of seronegative unvaccinated equines sentinel (28 animals: 10% of prevalence, 95% confidence level) was selected in order to uniformly cover the study area. Horses were sampled three times during the transmission season: immediately before (month of May), at seasonal peak (last week of August) and at the end of the transmission season (last week of September), with the aim to monitor the raising of WNV-specific IgM antibodies that are related to recent infection by commercial competitive ELISA. Positive samples were confirmed by seroneutralization assay (SN) and IgM ELISA at the CESME. Seroconversion was confirmed if SN titre was at least 1:10 and there was evidence of IgM antibodies [49].

Since 2014, the sentinel horses system in 'Garzaia di Marengo' was replaced by a one-shot random sampling of horses in order to detect IgM antibodies suggestive of WNV early infection. The number of animals to be tested for each Province during the transmission season was assigned

Since 2015, after the detection of WNV circulation in mosquito pools and in horses in Piemonte Region, veterinary surveillance procedures were revised, and the Region has been considered

Serological testing on horses was suspended while active virological surveillance on synanthropic non-migratory birds was implemented. From April to November, synanthropic birds (magpies—*Pica pica* and hooded crows—*Corvus cornix*), in the framework of the Regional control programme against pest birds for cultivated land protection, were weekly captured by Larsen traps and then killed or directly shot by trained hunters. Samples of organs (brain,

All samples were analysed by a TaqMan® One-Step RT-PCR protocol distinctive for WNV Lineage 1 and Lineage 2 with WN-LCV-F1 and WN-LCV-R1 primers [44] and a TaqMan®

In 2017, the control programme against synanthropic birds was not activated in Torino Province. Then, as alternatively recommended by the National Surveillance Plan, a specific serological monitoring activity was enforced on rural poultry, with the aim to detect IgG

Since 2014, Liguria Region was included in WNV National Surveillance plan by serological random sampling of horses in order to detect IgM antibodies suggestive of WNV early infection. The number of animals to be tested for each Province during the transmission season was assigned based on the provincial equine census, as recorded in National Livestock

**3.2. Human surveillance West Nile virus and others MBDs in Piemonte region and** 

Human surveillance activities against MBDs in Piemonte Region are regulated by both the National Surveillance Plan of imported and autochthonous MBDs (CHIKV, DENV and ZIKV), annually published by MoH and by National Integrated Surveillance Programme

based on the provincial equine census, as recorded in National Livestock Database [50].

as WNV Endemic Area by the National Surveillance programme.

66 Current Topics in Tropical Emerging Diseases and Travel Medicine

heart and kidney) of each bird were pooled and tested for WNV and USUV.

One-Step RT-PCR protocol for USUV with USU F and USU R primers [45].

antibodies by commercial ELISA test.

*3.1.4.2. Liguria region*

Database [50].

**Liguria region**

against WNV and USUV.
