o:36643 Izolacija i genska karakterizacija virusa afričke kuge svinja u Srbiji u periodu 2019-2023. godine doktorska disertacija Isolation and genetic characterisation of african swine fever virus in Serbia in the period 2019 - 2023 : doctoral dissertation sr Ova studija predstavlja prvi sveobuhvatan izveštaj o molekularnim karakteristikama sojeva virusa afričke kuge svinja (AKS) u Srbiji u periodu od 2019. do 2023. godine. Od prvog izbijanja bolesti u julu 2019. godine, bolest je registrovana kod divljih i domaćih svinja što je dovelo do smanjenja broja svinja u Srbiji za više od 19,7 %. U ovoj studiji, izvršena je molekularna analiza 95 sojeva virusa afričke kuge svinja iz 12 upravnih okruga u kojima je potvrđena bolest. Sekvenciranje je obuhvatilo četiri genska markera: B646L, E183L, B602L, i intergensku regiju (IGR) između gena I73R-I329L. Pored toga, izvršeno je i sekvenciranje celog genoma pet sojeva uključujući one koji su izazvali pojavu bolesti 2019. godine. Na primarnim kulturama tkiva poreklom od alveolarnih makrofaga i leukocita svinja, izvršena je izolacija virusa i to na primarnim leukocitima je uspešnost izolacije iznosila 72,7 %, dok je na alveolarnim makrofagama svinja iznosila 91,3 %. Prostorno-vremenskom analizom pojave AKS kod divljih svinja u periodu od 2020. do 2023. godine otkrivenoje postojanje pet klastera, dva u periodu 2020-2021, jedan u 2022, i dva u 2023. godini. Svi analizirani sojevi virusa AKS iz perioda 2019-2023. pripadaju genotipu II. Na osnovu analize nukleotidnih sekvenci srpskih sojeva utvrđeno je postojanje više podgrupa, kao i dve linije na osnovu CVR B602L gena od kojih je jedna identična sa referentnim sojem iz Gruzije, a druga sa mutacijom T318A. Takođe, po prvi put u Srbiji je identifikovana dominantna IGR II, kao i IGR III linija. Sekvenciranjem celih genoma virusa AKS ustanovljene su značajne mutacije u genima NP1450L, ECO2, O174L, K145R, MGF, grupi gena ACD, kao i delecija u genu L60L. Svi izolovani virusi su pokazali efekat hemadsorpcije i klasifikovani su u istu serogrupu kao i referentni soj Georgia 2007/1. Retrospektivnom analizom utvrđeno je da su dva nezavisna izbijanja AKS u 2019. godini proistekla iz različitih izvora, što dodatno ukazuje na složenost situacije i potrebu za nastavkom istraživanja radi boljeg razumevanja i kontrole širenja bolesti. This study presents the first comprehensive report on the molecular characteristics of African Swine Fever Virus (ASFV) strains in Serbia during the period from 2019 to 2023. Since its first occurence in July 2019, the disease has been detected in both wild and domestic pigs, leading to a reduction of the pig population in Serbia by more than 19,7 %. The study involved molecular analysis of 95 positive samples collected from 12 infected administrative districts. Sequencing covered four genetic markers: B646L, E183L, B602L, and the intergenic region (IGR) between the I73R and I329L genes. Additionally, whole-genome sequencing of five strains, including those responsible for the outbreak in 2019, was conducted. Virus isolation was performed on primary tissue cultures originating from alveolar macrophages and leukocytes of pigs, with successful isolation with 71.7 % on primary leukocytes and 92.3 % on primary alveolar macrophages assessed based on the hemadsorption effect. Spatial-temporal analysis of ASFV in wild pigs during the research period from 2020 to 2023 revealed the existence of five clusters, with two in the period 2020-2021, one in 2022, and two in 2023. All analyzed ASFV strains from the period 2019-2023 belong to genotype II. Nucleotide sequence analysis of Serbian strains revealed the existence of multiple subgroups, including two lines of the CVR B602L gene, one identical to the reference strain from Georgia, and another with the T318A mutation. The dominant IGR II line and the IGR III line, identified for the first time in Serbia, were also observed. Whole-genome sequencing of ASFV identified significant mutations in the NP1450L, ECO2, O174L, K145R, MGF, ACD gene group, as well as a deletion in the L60L gene, in addition to 10 insignificant mutations. All isolates demonstrated the hemadsorption effect and were classified in the same serogroup as the reference strain Georgia 2007/1. Retrospective analysis confirmed that two independent ASF outbreaks in 2019 originated from different sources, highlighting the complexity of the situation and the need for continued research to better understand and control the spread of the disease. Veterinarska medicina - / Veterinary medicine Datum odbrane: 27.06.2025. Afrička kuga svinja, Srbija, molekularna karakterizacija, genotip II, B646L gen, E183L gen, IGR region, hemadsorpcijski efekat, genomske mutacije, epizootiološki klasteri African Swine Fever, Serbia, Molecular Characterization, Genotype II, B646L gene, E183L gene, IGR region, Hemadsorption Effect, Genomic Mutations, Epizooziological Clusters 636.4.09:578.5(043.3) 91552100 177055241 91552101 9865 2025-10-06T11:43:23.053Z 45 no 46 Dimitrije, 1994- Glišić 73641225 2025 63 mentor Sonja, 1963- Radojičić 12637543 2025 63 mentor Vesna, 1980- Milićević 16675687 2025 63 član komisije Dejan, 1964- Krnjaić 13843815 2025 63 član komisije Radiša, 1981- Prodanović 16643431 2025 63 član komisije Ivan Toplak 129559305 2025 86 str. 3911450 http://phaidrabg.bg.ac.rs/o:36643 no yes 12 2025-10-06T11:43:23.320Z 70 11 1067068 1067081 1067083 Afrička kuga svinja, Srbija, molekularna karakterizacija, genotip II, B646L gen, E183L gen, IGR region, hemadsorpcijski efekat, genomske mutacije, epizootiološki klasteri African Swine Fever, Serbia, Molecular Characterization, Genotype II, B646L gene, E183L gene, IGR region, Hemadsorption Effect, Genomic Mutations, Epizooziological Clusters 636.4.09:578.5(043.3) 1738 2025