Objective: to study the level of the immune layer of the population to pathogens of natural focal infectious diseases in order to establish the epidemic activity of natural foci of particularly dangerous infectious diseases in the Rostov region.Materials and methods: blood serums of healthy donors living in the administrative territories of the region were collected in the period from 2020 to 2022. Specific antibodies in blood sera were determined by the ELISA method. Serum specific IgG class immunoglobulins were used as a serological marker of the transmitted infection.Results: IgG to the Crimean hemorrhagic fever virus was not detected in 2020. In 2021, the share of seropositive results was 0.7%, in 2022 — 2.0%. The proportion of IgG to West Nile virus in 2020 was 8.7%, in 2021 — 9.9%, in 2022 — 12.4%. Serological testing revealed an immune layer to ixodic tick–borne borreliosis in 2020–2022: 3.1%, 2.6% and 2.0%, respectively. Antibodies to hantaviruses, pathogens of hemorrhagic fever with renal syndrome in the blood of residents of the region in 2020 were found in 7.0% of samples, in 2021 — 4.5%, in 2022 — 7.1%. The share of positive samples in the study for Ku fever in 2022 was 2.3%. Conclusion: Seropositive samples were detected for all studied natural focal infections. The greatest variety of natural focal infections confirmed by serological monitoring data was detected in cities (Rostov-on-Don, Taganrog, Zernograd, Kamensk-Shakhtinsky) and districts of RO (Salsky, Neklinovsky, Remontnensky).Conclusion. The results of the serological monitoring made it possible to identify the circulation of pathogens of KGL, LZN, ICB, Ku fever, hantaviruses in the territory of two districts of the region. The detection of specific antibodies in the blood sera of healthy donors indicates the epidemic activity of natural foci. Also, a comprehensive study of healthy donors expanded the understanding of the area of the most relevant natural focal infections such as (KGL, LZN, ICB), and less frequently registered at present (HFRS and Ku fever).