Abstract
This study provides the first comprehensive report on the molecular characteristics of African swine fever virus (ASFV) variants in Serbia between 2019 and 2022. Since its first observation in July 2019, the disease has been found in wild boar and domestic swine. The study involved the analysis of 95 ASFV-positive samples collected from 12 infected administrative districts in Serbia. Partial four genomic regions were genetically characterized, including B646L, E183L, B602L, and the intergenic region (IGR) between the I73R-I329L genes. The results of the study suggest that multiple ASFV strains belonging to genotype II are circulating in Serbia, as evidenced by the analysis of the IGR between I73R-I329L genes that showed the most differences. Furthermore, the phylogenetic analysis of the B602L gene showed three different clades within the CVR I group of ASFV strains. Regarding the IGR, 98.4% were grouped into IGR II, with only one positive sample grouped into the IGR III group. These findings provide essential insights into the molecular characteristics of ASFV variants in Serbia and contribute to the knowledge of circulating strains of ASFV in Europe. However, further research is necessary to gain a better understanding of ASFV spread and evolution.
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Data availability
The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.
References
Achenbach JE, Gallardo C, Nieto-Pelegrín E et al (2017) Identification of a New Genotype of African Swine Fever Virus in Domestic Pigs from Ethiopia. Transbound Emerg Dis 64:1393–1404. https://doi.org/10.1111/tbed.12511
Alonso C, Borca M, Dixon L et al (2018) ICTV Virus Taxonomy Profile: Asfarviridae. J Gen Virol 99:613–614. https://doi.org/10.1099/jgv.0.001049
Bao J, Zhang Y, Shi C et al (2022) Genome-Wide Diversity Analysis of African Swine Fever Virus Based on a Curated Dataset. Animals 12:2446. https://doi.org/10.3390/ani12182446
Bastos ADS, Penrith M-L, Crucière et al (2003) Genotyping field strains of African swine fever virus by partial p72 gene characterisation. Arch Virol 148:693–706
Biek R, Pybus OG, Lloyd-Smith JO, Didelot X (2015) Measurably evolving pathogens in the genomic era. Trends Ecol Evol 30:306–313. https://doi.org/10.1016/j.tree.2015.03.009
Boshoff CI, Bastos ADS, Gerber LJ, Vosloo W (2007) Genetic characterisation of African swine fever viruses from outbreaks in southern Africa (1973–1999). Vet Microbiol 121:45–55. https://doi.org/10.1016/j.vetmic.2006.11.007
Cadenas-Fernández E, Ito S, Aguilar-Vega C et al (2022) The Role of the Wild Boar Spreading African Swine Fever Virus in Asia: Another Underestimated Problem. Front Vet Sci 9:453. https://doi.org/10.3389/FVETS.2022.844209/BIBTEX
Chenais E, Depner K, Guberti V et al (2019) Epidemiological considerations on African swine fever in Europe 2014–2018. Porcine Health Manag 5:6. https://doi.org/10.1186/s40813-018-0109-2
de la Vega I, González A, Blasco R et al (1994) Nucleotide Sequence and Variability of the Inverted Terminal Repetitions of African Swine Fever Virus DNA. Virology 201:152–156. https://doi.org/10.1006/viro.1994.1277
de Oliveira RP, Hutet E, Paboeuf F, et al (2019) Comparative vector competence of the Afrotropical soft tick Ornithodoros moubata and Palearctic species, O. erraticus and O. verrucosus, for African swine fever virus strains circulating in Eurasia. PLoS One 14:e0225657 https://doi.org/10.1371/JOURNAL.PONE.0225657
Denstedt E, Porco A, Hwang J et al (2021) Detection of African swine fever virus in free-ranging wild boar in Southeast Asia. Transbound Emerg Dis 68:2669–2675. https://doi.org/10.1111/TBED.13964
Department for Environment F and RAA and PHAAH and WA-IDM (2023) Updated Outbreak Assessment #24 - African swine fever (ASF) in Asia and Oceania. London
Dixon LK, Chapman DAG, Netherton CL, Upton C (2013) African swine fever virus replication and genomics. Virus Res 173:3–14. https://doi.org/10.1016/j.virusres.2012.10.020
European Centre for Disease Prevention and Control (ECDC) (2018) Ornithodoros spp. - current known distribution, May 2018
European Commission: Animal Disease Information System (2019) ADIS 2019
European Commission: Animal Disease Information System (2020) ADIS 2020
European Commission: Animal Disease Information System (2021) ADIS 2021
European Commission: Animal Disease Information System (2022) ADIS 2022
Faburay B (2022) Genome Plasticity of African Swine Fever Virus: Implications for Diagnostics and Live-Attenuated Vaccines. Pathogens 11:145. https://doi.org/10.3390/pathogens11020145
Forth JH, Calvelage S, Fischer M et al (2023) African swine fever virus – variants on the rise. Emerg Microbes Infect 12:2146537. https://doi.org/10.1080/22221751.2022.2146537
Forth JH, Forth LF, Blome S, et al (2020) African swine fever whole-genome sequencing—Quantity wanted but quality needed. PLoS Pathog 16:e1008779 https://doi.org/10.1371/journal.ppat.1008779
Gallardo C, Casado N, Soler A et al (2023) A multi gene-approach genotyping method identifies 24 genetic clusters within the genotype II-European African swine fever viruses circulating from 2007 to 2022. Front Vet Sci 10:1112850. https://doi.org/10.3389/fvets.2023.1112850
Gallardo C, Fernández-Pinero J, Pelayo V et al (2014) Genetic Variation among African Swine Fever Genotype II Viruses, Eastern and Central Europe. Emerg Infect Dis 20:1544–1547. https://doi.org/10.3201/eid2009.140554
Gallardo C, Mwaengo DM, Macharia JM et al (2009) Enhanced discrimination of African swine fever virus isolates through nucleotide sequencing of the p54, p72, and pB602L (CVR) genes. Virus Genes 38:85–95. https://doi.org/10.1007/s11262-008-0293-2
Gallardo C, Nurmoja I, Soler A et al (2018) Evolution in Europe of African swine fever genotype II viruses from highly to moderately virulent. Vet Microbiol 219:70–79. https://doi.org/10.1016/J.VETMIC.2018.04.001
Glišić D, Milićević V, Veljović L et al (2023) Patterns of ASFV Transmission in Domestic Pigs in Serbia. Pathogens 12:149. https://doi.org/10.3390/pathogens12010149
Gogin A, Gerasimov V, Malogolovkin A, Kolbasov D (2013) African swine fever in the North Caucasus region and the Russian Federation in years 2007–2012. Virus Res 173:198–203. https://doi.org/10.1016/j.virusres.2012.12.007
Iscaro C, Dondo A, Ruocco L et al (2022) Index case of new African Swine Fever incursion in mainland Italy. Transbound Emerg Dis. https://doi.org/10.1111/tbed.14584
Jean-Pierre RP, Hagerman AD, Rich KM (2022) An analysis of African Swine Fever consequences on rural economies and smallholder swine producers in Haiti. Front Vet Sci 9:1293. https://doi.org/10.3389/FVETS.2022.960344/BIBTEX
Jo YS, Gortázar C (2021) African Swine Fever in wild boar: Assessing interventions in South Korea. Transbound Emerg Dis 68:2878–2889. https://doi.org/10.1111/TBED.14106
Kim S, Lee S, Jeong H et al (2021) Rapid emergence of African swine fever virus variants with different numbers of a tandem repeat sequence in South Korea. Transbound Emerg Dis 68:1726–1730. https://doi.org/10.1111/tbed.13867
King DP, Reid SM, Hutchings GH et al (2003) Development of a TaqMan® PCR assay with internal amplification control for the detection of African swine fever virus. J Virol Methods 107:53–61. https://doi.org/10.1016/S0166-0934(02)00189-1
Koeltz A, Kolbasov D, Titov I, et al (2018) African Swine Fever Virus, Siberia, Russia, 2017. Emerg Infect Dis • www.cdc.gov/eid • 24 https://doi.org/10.1016/j.idcr.2014.12.002
Ladoși I, Păpuc TA, Ladoși D (2023) The impact of african swine fever (ASF) on Romanian pig meat production: A Review. Acta Vet-Bel 73:1–12. https://doi.org/10.2478/acve-2023-0001
Li X, Xiao K, Zhang Z et al (2020) The recombination hot spots and genetic diversity of the genomes of African swine fever viruses. J Infect 80:121–142. https://doi.org/10.1016/j.jinf.2019.08.007
Mazloum A, van Schalkwyk A, Chernyshev R et al (2023a) A Guide to Molecular Characterization of Genotype II African Swine Fever Virus: Essential and Alternative Genome Markers. Microorganisms 11:642. https://doi.org/10.3390/MICROORGANISMS11030642/S1
Mazloum A, Van Schalkwyk A, Chernyshev R et al (2022) Genetic Characterization of the Central Variable Region in African Swine Fever Virus Isolates in the Russian Federation from 2013 to 2017. Pathogens 11:919. https://doi.org/10.3390/pathogens11080919
Mazloum A, van Schalkwyk A, Shotin A et al (2023b) Whole-genome sequencing of African swine fever virus from wild boars in the Kaliningrad region reveals unique and distinguishing genomic mutations. Front Vet Sci 9:1976. https://doi.org/10.3389/FVETS.2022.1019808/BIBTEX
Mazur-Panasiuk N, Walczak M, Juszkiewicz M, Woźniakowski G (2020) The Spillover of African Swine Fever in Western Poland Revealed Its Estimated Origin on the Basis of O174L, K145R, MGF 505–5R and IGR I73R/I329L Genomic Sequences. Viruses 12:1094. https://doi.org/10.3390/v12101094
Mazur-Panasiuk N, Woźniakowski G, Niemczuk K (2019) The first complete genomic sequences of African swine fever virus isolated in Poland. Sci Rep 9:4556. https://doi.org/10.1038/s41598-018-36823-0
Milićević V, Kureljušić B, Maksimović Zorić J et al (2019) First Occurence of African Swine Fever in Serbia. Acta Vet Brno 69:443–449. https://doi.org/10.2478/acve-2019-0038
Nešković M, Ristić B, Došenović R et al (2021) African Swine Fever Outbreak Investigation on Large Commercial Pig Farm in Serbia. Acta Vet Brno 71:219–229. https://doi.org/10.2478/acve-2021-0019
Nguyen VT, Cho K, Mai NTA et al (2022) Multiple variants of African swine fever virus circulating in Vietnam. Arch Virol 167:1137–1140. https://doi.org/10.1007/s00705-022-05363-4
Nix RJ, Gallardo C, Hutchings G et al (2006) Molecular epidemiology of African swine fever virus studied by analysis of four variable genome regions. Arch Virol 151:2475–2494. https://doi.org/10.1007/s00705-006-0794-z
Nurmoja I, Petrov A, Breidenstein C et al (2017) Biological characterization of African swine fever virus genotype II strains from North-Eastern Estonia in European wild boar. Transbound Emerg Dis 64:2034–2041. https://doi.org/10.1111/TBED.12614
Oļševskis E, Guberti V, Seržants M et al (2016) African swine fever virus introduction into the EU in 2014: Experience of Latvia. Res Vet Sci 105:28–30. https://doi.org/10.1016/J.RVSC.2016.01.006
Pereira De Oliveira R, Hutet E, Lancelot R et al (2020) Differential vector competence of Ornithodoros soft ticks for African swine fever virus: What if it involves more than just crossing organic barriers in ticks? Parasit Vectors 13:1–15. https://doi.org/10.1186/S13071-020-04497-1/FIGURES/6
Pikalo J, Schoder M, Sehl J et al (2020) The African swine fever virus isolate Belgium 2018/1 shows high virulence in European wild boar. Transbound Emerg Dis 67:1654–1659. https://doi.org/10.1111/tbed.13503
Polaček V, Mirčeta J, Prodanov-Radulović J (2021) Key risk factors and impact of African swine fever spreading on pig production in Serbia. Acta Vet Bel 71:371–391. https://doi.org/10.2478/acve-2021-0032
Prodanov-Radulović J, Mirčeta J, Djurdjević B, et al (2023) African Swine Fever Outbreak in an Enclosed Wild Boar Hunting Ground in Serbia. Pathogens 12:691https://doi.org/10.3390/PATHOGENS12050691
Quembo CJ, Jori F, Vosloo W, Heath L (2017) Genetic characterization of African swine fever virus isolates from soft ticks at the wildlife/domestic interface in Mozambique and identification of a novel genotype. https://doi.org/10.1111/tbed.12700
Quembo CJ, Jori F, Heath L et al (2016) Investigation into the Epidemiology of African Swine Fever Virus at the Wildlife - Domestic Interface of the Gorongosa National Park, Central Mozambique. Transbound Emerg Dis 63:443–451. https://doi.org/10.1111/tbed.12289
Ramishvili Levan (2007) African swine fever - Georgia: OIE. In: ProMED INTERNATIONAL SOCIETY FOR INFECTIOUS DISEASES. https://promedmail.org/promed-post/?id=12219. Accessed 27 Mar 2023
Rowlands RJ, Michaud V, Heath L et al (2008) African Swine Fever Virus Isolate, Georgia, 2007. Emerg Infect Dis 14:1870–1874. https://doi.org/10.3201/eid1412.080591
Sargsyan MA, Voskanyan HE, Karalova EM, et al (2018) Third wave of African swine fever infection in Armenia: Virus demonstrates the reduction of pathogenicity. Vet World https://doi.org/10.14202/vetworld.2018.5-9
Sauter-Louis C, Conraths FJ, Probst C et al (2021a) African Swine Fever in Wild Boar in Europe—A Review. Viruses 13:1717. https://doi.org/10.3390/v13091717
Sauter-Louis C, Forth JH, Probst C et al (2021b) Joining the club: First detection of African swine fever in wild boar in Germany. Transbound Emerg Dis 68:1744–1752. https://doi.org/10.1111/TBED.13890
Shi K, Liu H, Yin Y, et al (2022) Molecular Characterization of African Swine Fever Virus From 2019–2020 Outbreaks in Guangxi Province, Southern China. Front Vet Sci 9:912224 https://doi.org/10.3389/fvets.2022.912224
Sun E, Huang L, Zhang X et al (2021) Genotype I African swine fever viruses emerged in domestic pigs in China and caused chronic infection. Emerg Microbes Infect 10:2183–2193. https://doi.org/10.1080/22221751.2021.1999779
Tran HTT, Truong AD, Dang AK et al (2021) Circulation of two different variants of intergenic region (IGR) located between the I73R and I329L genes of African swine fever virus strains in Vietnam. Transbound Emerg Dis 68:2693–2695. https://doi.org/10.1111/tbed.13996
Vilem A, Nurmoja I, Niine T et al (2020) Molecular Characterization of African Swine Fever Virus Isolates in Estonia in 2014–2019. Pathogens 9:582. https://doi.org/10.3390/pathogens9070582
WOAH (2022) WAHIS. https://wahis.woah.org/#/in-event/4761/dashboard. Accessed 27 Mar 2023
Woźniakowski G, Pejsak Z, Jabłoński A (2021) Emergence of African Swine Fever in Poland (2014–2021). Successes and Failures in Disease Eradication. Agriculture 11:738https://doi.org/10.3390/AGRICULTURE11080738
Acknowledgements
The sequences were generated through the Sequencing Service of the Animal Production and Health Sub-Programme of the Joint FAO/IAEA Division in Vienna, Austria.
Funding
The Serbian Ministry of Science, Technological Development and Innovation provided funding for the study through Contract No 451–03-47/2023–01/200030 and 451–03-47/2023–01/200143.
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All authors contributed to the study conception and design. D.G. was the principal investigator, molecular tests were carried out by D.G. and V.M., sequences were analysed by I.T. and C.G., the epidemiological investigation was carried out by R.P. and S.R., and D.K. and S.R. supervised the study.
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Glišić, D., Milićević, V., Krnjaić, D. et al. Genetic analysis reveals multiple intergenic region and central variable region in the African swine fever virus variants circulating in Serbia. Vet Res Commun 47, 1925–1936 (2023). https://doi.org/10.1007/s11259-023-10145-7
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DOI: https://doi.org/10.1007/s11259-023-10145-7