Serological monitoring of backyard chickens in Central Macedonia-Greece can detect low transmission of West Nile virus in the absence of human neuroinvasive disease cases

During 2010-13, West Nile virus (WNV) epidemics occurred in Greece with high numbers of human cases. In parallel, WNV serological surveillance utilizing domestic birds was applied mainly in Central Macedonia, as well as in other areas of the country, and allowed efficient detection of WNV activity during this period. The objective of the study was to evaluate the sensitivity of chicken-based WNV surveillance in periods of low-level virus transmission (2014-15) in a well-studied area, i.e. the epicenter of the 2010 WNV epidemic (Central Macedonia), which is considered endemic since then. WNV activity was monitored via determination of antiviral immune responses in juvenile backyard chickens. The birds were sampled twice per transmission period. WNV-specific antibodies were detected by ELISA in 2.8% out of 255 chickens sampled early in the 2014 transmission period (95% CI: 1-6%). Continued virus transmission was detected at the end of the period, as 4.2% out of 240 sampled chickens seroconverted to WNV (95% CI: 2-8%). Although 14 human neuroinvasive cases occurred in Greece during 2014, no such cases were reported in the study area. During the 2015 early warning period, antibodies against WNV were not detected in sampled chickens (n = 250, 95% CI: 0-2%). However, humoral immune responses were detected in 6 out of 240 chicken sampled at the end of the transmission period (2.5%; 95% CI: 1-6%), indicating continued WNV activity. No human cases were reported in Greece during 2015. All samples were negative with real-time RT-PCR. Serological surveillance of chickens resulted in identification of areas with low WNV activity levels during 2014-15, and provided indications of its overwintering in Central Macedonia. The findings suggest that surveillance based on serological testing of domestic birds is sensitive and able to detect low-level of WNV enzootic transmission, in the absence of human cases.