In Serbia, the West Nile virus (WNV) has been a persistent threat for the past ten years. From 2012-2022, epidemics of varying sizes have been reported among the population in Serbia. The annual national WNV monitoring program in Serbia was started by veterinary services in June 2022, as a continuation of the surveillance program started in 2014. The Veterinary Directorate funded the program, which was carried out by veterinary services with entomologists and ornithologists. The monitoring program's primary goal was the early discovery of WNV in specific areas and prompt notification of authorities, human health agencies and the public. The surveillance program is based upon direct and indirect detection of WNV in different areas. Serological tests of horses for the presence of anti-WNV-IgM antibodies and sentinel calves aged 6-8 months (calved after the vector season in the previous year) for the presence of anti-WNV-IgG antibodies were used to indirectly monitor the virus's presence and confirm infection. Direct monitoring of WNV presence was carried out through molecular testing of wild birds and pooled mosquito samples under active surveillance and through testing of clinically suspect horses, under passive surveillance. Total number of samples per district was determined according to WNV presence and infection risk. From June-September, 2022, 1853 blood samples of horses were examined and seroconversion (IgM antibodies) was determined in 25(1.3%) of tested horses. Positive serological responses were found in 0.9%(4/437), 2.0%(10/490), 1.9%(9/468), and 0.9%(4/458) of examined horses in June, July, August, and September, respectively. The seroconversion occurred in an upward line in accordance with vector activity increase. There were 2 districts in June, 3 in July, 5 in August, and 3 in September in which seropositive horses were found. The presence of anti-WNV-IgG antibodies was found in 8.0% (140/1740) of sentinel calves, during the same time period in high-risk districts (14/25), specifically in 11(2.7%), 34(6.9%), 46(10.6%), and 49(11.8%) tested sentinel calves in 4/14, 7/14, 9/14, and 9/14 districts in June, July, August, and September, respectively. From June-September 2022, 792 pooled mosquito samples were analysed for WNV presence and 34(4.3%) of the samples tested positive. From the first negative result in June (0/160) to the first positive discoveries in July 5.9% (19/320), continuing with positive findings in August 4.5%(7/156), and finally in September 5.1%(8/156), the prevalence of WNV was in an upward trajectory. Nine out of Serbia's twenty five districts had positive mosquito sample findings: >20% of the tested samples were positive in the Belgrade region, 19.15% in Central Banat, 18.37% in South Banat, 8.7% in Srem, 8.33% in South Ba?ka, 6.0% in Šumadia, 4.17% in North Banat, 4.08% in North Ba?ka, and 2.0% in West Ba?ka district. WNV was identified in 14(3.7%) out of 379 analyzed pharyngeal swab samples of live captured wild birds in four districts and in 11 samples out of 52(21.1%) dead wild birds in seven districts. To maintain a prompt response and protection of human and animal health, it is required to sustain and strengthen the WNV monitoring program that was implemented throughout 2022.