The first report of porcine parvovirus 7 (PPV7) in Colombia demonstrates the presence of variants associated with modifications at the level of the VP2-capsid protein
There are a wide variety of porcine parvoviruses (PPVs) referred to as PPV1 to PPV7. The latter was discovered in 2016 and later reported in some countries in America, Asia, and Europe. PPV7 as a pathogenic agent or coinfection with other pathogens causing disease has not yet been determined. In the...
- Autores:
-
Vargas Bermúdez, Diana
Rendón Marín, Santiago
Ruiz Sáenz, Julián
Mogollón, José
Jaime, Jairo
- Tipo de recurso:
- Article of investigation
- Fecha de publicación:
- 2021
- Institución:
- Universidad Cooperativa de Colombia
- Repositorio:
- Repositorio UCC
- Idioma:
- OAI Identifier:
- oai:repository.ucc.edu.co:20.500.12494/43972
- Acceso en línea:
- https://hdl.handle.net/20.500.12494/43972
- Palabra clave:
- Capsids
Swine
Phylogenetic analysis
B cells
Nucleotide sequencing
Parvoviruses
Colombia
Structural proteins
- Rights
- openAccess
- License
- Atribución – No comercial – Compartir igual
| id |
COOPER2_c5982d2c312dd1d35b4a8905cf93d777 |
|---|---|
| oai_identifier_str |
oai:repository.ucc.edu.co:20.500.12494/43972 |
| network_acronym_str |
COOPER2 |
| network_name_str |
Repositorio UCC |
| repository_id_str |
|
| dc.title.spa.fl_str_mv |
The first report of porcine parvovirus 7 (PPV7) in Colombia demonstrates the presence of variants associated with modifications at the level of the VP2-capsid protein |
| title |
The first report of porcine parvovirus 7 (PPV7) in Colombia demonstrates the presence of variants associated with modifications at the level of the VP2-capsid protein |
| spellingShingle |
The first report of porcine parvovirus 7 (PPV7) in Colombia demonstrates the presence of variants associated with modifications at the level of the VP2-capsid protein Capsids Swine Phylogenetic analysis B cells Nucleotide sequencing Parvoviruses Colombia Structural proteins |
| title_short |
The first report of porcine parvovirus 7 (PPV7) in Colombia demonstrates the presence of variants associated with modifications at the level of the VP2-capsid protein |
| title_full |
The first report of porcine parvovirus 7 (PPV7) in Colombia demonstrates the presence of variants associated with modifications at the level of the VP2-capsid protein |
| title_fullStr |
The first report of porcine parvovirus 7 (PPV7) in Colombia demonstrates the presence of variants associated with modifications at the level of the VP2-capsid protein |
| title_full_unstemmed |
The first report of porcine parvovirus 7 (PPV7) in Colombia demonstrates the presence of variants associated with modifications at the level of the VP2-capsid protein |
| title_sort |
The first report of porcine parvovirus 7 (PPV7) in Colombia demonstrates the presence of variants associated with modifications at the level of the VP2-capsid protein |
| dc.creator.fl_str_mv |
Vargas Bermúdez, Diana Rendón Marín, Santiago Ruiz Sáenz, Julián Mogollón, José Jaime, Jairo |
| dc.contributor.author.none.fl_str_mv |
Vargas Bermúdez, Diana Rendón Marín, Santiago Ruiz Sáenz, Julián Mogollón, José Jaime, Jairo |
| dc.subject.spa.fl_str_mv |
Capsids Swine Phylogenetic analysis B cells Nucleotide sequencing Parvoviruses Colombia Structural proteins |
| topic |
Capsids Swine Phylogenetic analysis B cells Nucleotide sequencing Parvoviruses Colombia Structural proteins |
| description |
There are a wide variety of porcine parvoviruses (PPVs) referred to as PPV1 to PPV7. The latter was discovered in 2016 and later reported in some countries in America, Asia, and Europe. PPV7 as a pathogenic agent or coinfection with other pathogens causing disease has not yet been determined. In the present study, we report the identification of PPV7 for the first time in Colombia, where it was found retrospectively since 2015 in 40% of the provinces that make up the country (13/32), and the virus was ratified for 2018 in 4/5 provinces evaluated. Additionally, partial sequencing (nucleotides 380 to 4000) was performed of four Colombian strains completely covering the VP2 and NS1 viral genes. A sequence identity greater than 99% was found when comparing them with reference strains from the USA and China. In three of the four Colombian strains, an insertion of 15 nucleotides (five amino acids) was found in the PPV7-VP2 capsid protein (540–5554 nt; 180–184 aa). Based on this insertion, the VP2 phylogenetic analysis exhibited two well-differentiated evolutionarily related groups. To evaluate the impact of this insertion on the structure of the PPV7-VP2 capsid protein, the secondary structure of two different Colombian strains was predicted, and it was determined that the insertion is located in the coil region and not involved in significant changes in the structure of the protein. The 3D structure of the PPV7-VP2 capsid protein was determined by threading and homology modeling, and it was shown that the insertion did not imply a change in the shape of the protein. Additionally, it was determined that the insertion is not involved in suppressing a potential B cell epitope, although the increase in length of the epitope could affect the interaction with molecules that allow a specific immune response. |
| publishDate |
2021 |
| dc.date.issued.none.fl_str_mv |
2021-12 |
| dc.date.accessioned.none.fl_str_mv |
2022-02-15T20:20:49Z |
| dc.date.available.none.fl_str_mv |
2022-02-15T20:20:49Z |
| dc.type.none.fl_str_mv |
Artículos Científicos |
| dc.type.coar.none.fl_str_mv |
http://purl.org/coar/resource_type/c_2df8fbb1 |
| dc.type.coarversion.none.fl_str_mv |
http://purl.org/coar/version/c_970fb48d4fbd8a85 |
| dc.type.driver.none.fl_str_mv |
info:eu-repo/semantics/article |
| dc.type.version.none.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
| format |
http://purl.org/coar/resource_type/c_2df8fbb1 |
| status_str |
publishedVersion |
| dc.identifier.issn.spa.fl_str_mv |
1932-6203 |
| dc.identifier.uri.spa.fl_str_mv |
10.1371/journal.pone.0258311 |
| dc.identifier.uri.none.fl_str_mv |
https://hdl.handle.net/20.500.12494/43972 |
| dc.identifier.bibliographicCitation.spa.fl_str_mv |
Vargas-Bermudez DS, Rendon-Marin S, Ruiz-Saenz J, Mogollón D, Jaime J (2021) The first report of porcine parvovirus 7 (PPV7) in Colombia demonstrates the presence of variants associated with modifications at the level of the VP2-capsid protein. PLOS ONE 16(12): e0258311. https://doi.org/10.1371/journal.pone.0258311 |
| identifier_str_mv |
1932-6203 10.1371/journal.pone.0258311 Vargas-Bermudez DS, Rendon-Marin S, Ruiz-Saenz J, Mogollón D, Jaime J (2021) The first report of porcine parvovirus 7 (PPV7) in Colombia demonstrates the presence of variants associated with modifications at the level of the VP2-capsid protein. PLOS ONE 16(12): e0258311. https://doi.org/10.1371/journal.pone.0258311 |
| url |
https://hdl.handle.net/20.500.12494/43972 |
| dc.relation.isversionof.spa.fl_str_mv |
https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0258311 |
| dc.relation.ispartofjournal.spa.fl_str_mv |
PLOS ONE |
| dc.relation.references.spa.fl_str_mv |
Ranz AI, Manclús JJ, Díaz-Aroca E, Casal JI. Porcine parvovirus: DNA sequence and genome organization. J Gen Virol. 1989;70 (Pt 10): 2541–2553. pmid:2794971 Pénzes JJ, Söderlund-Venermo M, Canuti M, Eis-Hübinger AM, Hughes J, Cotmore SF, et al. Reorganizing the family Parvoviridae: a revised taxonomy independent of the canonical approach based on host association. Arch Virol. 2020; pmid:32533329 Yang S, Liu Z, Wang Y, Li W, Fu X, Lin Y, et al. A novel rodent Chapparvovirus in feces of wild rats. Virol J. 2016;13: 133. pmid:27473724 Williams SH, Che X, Garcia JA, Klena JD, Lee B, Muller D, et al. Viral diversity of house mice in new york city. MBio. 2018;9. pmid:29666290 Fahsbender E, Altan E, Seguin MA, Young P, Estrada M, Leutenegger C, et al. Chapparvovirus DNA Found in 4% of Dogs with Diarrhea. Viruses. 2019;11. pmid:31035625 Reuter G, Boros Á, Delwart E, Pankovics P. Novel circular single-stranded DNA virus from turkey faeces. Arch Virol. 2014;159: 2161–2164. pmid:24562429 Lima DA, Cibulski SP, Tochetto C, Varela APM, Finkler F, Teixeira TF, et al. The intestinal virome of malabsorption syndrome-affected and unaffected broilers through shotgun metagenomics. Virus Res. 2019;261: 9–20. pmid:30543873 Souza WM de Romeiro MF, Fumagalli MJ, Modha S, de Araujo J, Queiroz LH, et al. Chapparvoviruses occur in at least three vertebrate classes and have a broad biogeographic distribution. J Gen Virol. 2017;98: 225–229. pmid:28284244 Palinski RM, Mitra N, Hause BM. Discovery of a novel Parvovirinae virus, porcine parvovirus 7, by metagenomic sequencing of porcine rectal swabs. Virus Genes. 2016;52: 564–567. pmid:26995221 Xing X, Zhou H, Tong L, Chen Y, Sun Y, Wang H, et al. First identification of porcine parvovirus 7 in China. Arch Virol. 2018;163: 209–213. pmid:29022179 Wang Y, Yang K-K, Wang J, Wang X-P, Zhao L, Sun P, et al. Detection and molecular characterization of novel porcine parvovirus 7 in Anhui province from Central-Eastern China. Infect Genet Evol. 2019;71: 31–35. pmid:30876889 Wang W, Cao L, Sun W, Xin J, Zheng M, Tian M, et al. Sequence and phylogenetic analysis of novel porcine parvovirus 7 isolates from pigs in Guangxi, China. PLoS One. 2019;14: e0219560. pmid:31291362 Ouh I-O, Park S, Lee J-Y, Song JY, Cho I-S, Kim H-R, et al. First detection and genetic characterization of porcine parvovirus 7 from Korean domestic pig farms. J Vet Sci. 2018;19: 855–857. pmid:30304892 Chung H-C, Nguyen V-G, Huynh T-M-L, Park Y-H, Park K-T, Park B-K. PCR-based detection and genetic characterization of porcine parvoviruses in South Korea in 2018. BMC Vet Res. 2020;16: 113. pmid:32295585 Miłek D, Woźniak A, Stadejek T. The detection and genetic diversity of novel porcine parvovirus 7 (PPV7) on Polish pig farms. Res Vet Sci. 2018;120: 28–32. pmid:30170185 Blomström A-L, Ye X, Fossum C, Wallgren P, Berg M. Characterisation of the Virome of Tonsils from Conventional Pigs and from Specific Pathogen-Free Pigs. Viruses. 2018;10. pmid:30036964 Da Silva MS, Budaszewski RF, Weber MN, Cibulski SP, Paim WP, Mósena ACS, et al. Liver virome of healthy pigs reveals diverse small ssDNA viral genomes. Infect Genet Evol. 2020;81: 104203. pmid:32035977 Shackelton LA, Parrish CR, Truyen U, Holmes EC. High rate of viral evolution associated with the emergence of carnivore parvovirus. Proc Natl Acad Sci USA. 2005;102: 379–384. pmid:15626758 Streck AF, Canal CW, Truyen U. Molecular epidemiology and evolution of porcine parvoviruses. Infect Genet Evol. 2015;36: 300–306. pmid:26453771 Ren X, Tao Y, Cui J, Suo S, Cong Y, Tijssen P. Phylogeny and evolution of porcine parvovirus. Virus Res. 2013;178: 392–397. pmid:24050995 Wang D, Mai J, Yang Y, Wang N. Porcine Parvovirus 7: Evolutionary Dynamics and Identification of Epitopes toward Vaccine Design. Vaccines (Basel). 2020;8. pmid:32635618 Streck AF, Bonatto SL, Homeier T, Souza CK, Gonçalves KR, Gava D, et al. High rate of viral evolution in the capsid protein of porcine parvovirus. J Gen Virol. 2011;92: 2628–2636. pmid:21795474 Cadar D, Cságola A, Kiss T, Tuboly T. Capsid protein evolution and comparative phylogeny of novel porcine parvoviruses. Mol Phylogenet Evol. 2013;66: 243–253. pmid:23044400 Cotmore SF, Agbandje-McKenna M, Chiorini JA, Mukha DV, Pintel DJ, Qiu J, et al. The family Parvoviridae. Arch Virol. 2014;159: 1239–1247. pmid:24212889 Mengeling WL, Lager KM, Vorwald AC. The effect of porcine parvovirus and porcine reproductive and respiratory syndrome virus on porcine reproductive performance. Anim Reprod Sci. 2000;60–61: 199–210. pmid:10844195 Miłek D, Woźniak A, Podgórska K, Stadejek T. Do porcine parvoviruses 1 through 7 (PPV1-PPV7) have an impact on porcine circovirus type 2 (PCV2) viremia in pigs? Vet Microbiol. 2020;242: 108613. pmid:32122579 Opriessnig T, Fenaux M, Yu S, Evans RB, Cavanaugh D, Gallup JM, et al. Effect of porcine parvovirus vaccination on the development of PMWS in segregated early weaned pigs coinfected with type 2 porcine circovirus and porcine parvovirus. Vet Microbiol. 2004;98: 209–220. pmid:15036529 Allan GM, Kennedy S, McNeilly F, Foster JC, Ellis JA, Krakowka SJ, et al. Experimental reproduction of severe wasting disease by co-infection of pigs with porcine circovirus and porcine parvovirus. J Comp Pathol. 1999;121: 1–11. pmid:10373289 Ha Z, Xie C-Z, Li J-F, Wen S-B, Zhang K-L, Nan F-L, et al. Molecular detection and genomic characterization of porcine circovirus 3 in pigs from Northeast China. BMC Vet Res. 2018;14: 321. pmid:30367641 Oraveerakul K, Choi CS, Molitor TW. Tissue tropisms of porcine parvovirus in swine. Arch Virol. 1993;130: 377–389. pmid:8390826 Paul PS, Mengeling WL, Pirtle EC. Duration and biological half-life of passively acquired colostral antibodies to porcine parvovirus. Am J Vet Res. 1982;43: 1376–1379. pmid:7103222 Agbandje-McKenna M, Llamas-Saiz AL, Wang F, Tattersall P, Rossmann MG. Functional implications of the structure of the murine parvovirus, minute virus of mice. Structure. 1998;6: 1369–1381. pmid:9817841 Boisvert M, Fernandes S, Tijssen P. Multiple pathways involved in porcine parvovirus cellular entry and trafficking toward the nucleus. J Virol. 2010;84: 7782–7792. pmid:20484503 Simpson AA, Hébert B, Sullivan GM, Parrish CR, Zádori Z, Tijssen P, et al. The structure of porcine parvovirus: comparison with related viruses. J Mol Biol. 2002;315: 1189–1198. pmid:11827486 López-Bueno A, Rubio M-P, Bryant N, McKenna R, Agbandje-McKenna M, Almendral JM. Host-selected amino acid changes at the sialic acid binding pocket of the parvovirus capsid modulate cell binding affinity and determine virulence. J Virol. 2006;80: 1563–1573. pmid:16415031 Ji P, Liu Y, Chen Y, Wang A, Jiang D, Zhao B, et al. Porcine parvovirus capsid protein expressed in Escherichia coli self-assembles into virus-like particles with high immunogenicity in mice and guinea pigs. Antiviral Res. 2017;139: 146–152. pmid:28063996 Antonis AFG, Bruschke CJM, Rueda P, Maranga L, Casal JI, Vela C, et al. A novel recombinant virus-like particle vaccine for prevention of porcine parvovirus-induced reproductive failure. Vaccine. 2006;24: 5481–5490. pmid:16730104 Hueffer K, Parker JSL, Weichert WS, Geisel RE, Sgro J-Y, Parrish CR. The natural host range shift and subsequent evolution of canine parvovirus resulted from virus-specific binding to the canine transferrin receptor. J Virol. 2003;77: 1718–1726. pmid:12525605 Hu W, Xu X, Liu Q, Ji J, Kan Y, Yao L, et al. Molecular characterisation and genetic diversity of canine parvovirus type 2 prevalent in central china. J Vet Res. 2020;64: 347–354. pmid:32984622 Gainor K, Bowen A, Bolfa P, Peda A, Malik YS, Ghosh S. Molecular Investigation of Canine Parvovirus-2 (CPV-2) Outbreak in Nevis Island: Analysis of the Nearly Complete Genomes of CPV-2 Strains from the Caribbean Region. Viruses. 2021;13. pmid:34204082 Chen B, Zhang X, Zhu J, Liao L, Bao E. Molecular Epidemiological Survey of Canine Parvovirus Circulating in China from 2014 to 2019. Pathogens. 2021;10. pmid:34064982 Parrish CR. Mapping specific functions in the capsid structure of canine parvovirus and feline panleukopenia virus using infectious plasmid clones. Virology. 1991;183: 195–205. pmid:1647068 Kumar S, Stecher G, Tamura K. MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol Biol Evol. 2016;33: 1870–1874. pmid:27004904 Jones DT. Protein secondary structure prediction based on position-specific scoring matrices. J Mol Biol. 1999;292: 195–202. pmid:10493868 Roy A, Kucukural A, Zhang Y. I-TASSER: a unified platform for automated protein structure and function prediction. Nat Protoc. 2010;5: 725–738. pmid:20360767 Yang J, Yan R, Roy A, Xu D, Poisson J, Zhang Y. The I-TASSER Suite: protein structure and function prediction. Nat Methods. 2015;12: 7–8. pmid:25549265 Yang J, Zhang Y. I-TASSER server: new development for protein structure and function predictions. Nucleic Acids Res. 2015;43: W174–W181. pmid:25883148 Eswar N, Webb B, Marti-Renom MA, Madhusudhan MS, Eramian D, Shen M-Y, et al. Comparative protein structure modeling using Modeller. Curr Protoc Bioinformatics. 2006;Chapter 5: Unit 5.6. pmid:18428767 Xu D, Zhang Y. Improving the physical realism and structural accuracy of protein models by a two-step atomic-level energy minimization. Biophys J. 2011;101: 2525–2534. pmid:22098752 Wiederstein M, Sippl MJ. ProSA-web: interactive web service for the recognition of errors in three-dimensional structures of proteins. Nucleic Acids Res. 2007;35: W407–10. pmid:17517781 Sippl MJ. Recognition of errors in three-dimensional structures of proteins. Proteins. 1993;17: 355–362. pmid:8108378 Laskowski RA, MacArthur MW, Moss DS, Thornton JM. PROCHECK: a program to check the stereochemical quality of protein structures. J Appl Crystallogr. 1993;26: 283–291. Zhang Y, Skolnick J. TM-align: a protein structure alignment algorithm based on the TM-score. Nucleic Acids Res. 2005;33: 2302–2309. pmid:15849316 Bowie JU, Lüthy R, Eisenberg D. A method to identify protein sequences that fold into a known three-dimensional structure. Science. 1991;253: 164–170. pmid:1853201 Lüthy R, Bowie JU, Eisenberg D. Assessment of protein models with three-dimensional profiles. Nature. 1992;356: 83–85. pmid:1538787 Pettersen EF, Goddard TD, Huang CC, Couch GS, Greenblatt DM, Meng EC, et al. UCSF Chimera—a visualization system for exploratory research and analysis. J Comput Chem. 2004;25: 1605–1612. pmid:15264254 Jespersen MC, Peters B, Nielsen M, Marcatili P. BepiPred-2.0: improving sequence-based B-cell epitope prediction using conformational epitopes. Nucleic Acids Res. 2017;45: W24–W29. pmid:28472356 Yao B, Zhang L, Liang S, Zhang C. SVMTriP: a method to predict antigenic epitopes using support vector machine to integrate tri-peptide similarity and propensity. PLoS One. 2012;7: e45152. pmid:22984622 |
| dc.rights.license.none.fl_str_mv |
Atribución – No comercial – Compartir igual |
| dc.rights.accessrights.none.fl_str_mv |
info:eu-repo/semantics/openAccess |
| dc.rights.coar.none.fl_str_mv |
http://purl.org/coar/access_right/c_abf2 |
| rights_invalid_str_mv |
Atribución – No comercial – Compartir igual http://purl.org/coar/access_right/c_abf2 |
| eu_rights_str_mv |
openAccess |
| dc.format.extent.spa.fl_str_mv |
e0258311 |
| dc.coverage.temporal.spa.fl_str_mv |
16 |
| dc.publisher.spa.fl_str_mv |
Universidad Cooperativa de Colombia, Facultad de Ciencias de la Salud, Medicina Veterinaría y Zootecnia, Bucaramanga PLOS |
| dc.publisher.program.spa.fl_str_mv |
Medicina veterinaria y zootecnia |
| dc.publisher.place.spa.fl_str_mv |
Bucaramanga |
| institution |
Universidad Cooperativa de Colombia |
| bitstream.url.fl_str_mv |
https://repository.ucc.edu.co/bitstreams/c5a067e7-24ab-4ed6-8158-1ae42637e1dc/download https://repository.ucc.edu.co/bitstreams/8149ddcf-0f8b-4161-88d8-a01dfdc933e9/download https://repository.ucc.edu.co/bitstreams/8ca5238d-d9ba-4246-bae4-bc8ac9d3ea26/download https://repository.ucc.edu.co/bitstreams/93819280-1a47-4f10-9c26-f0966f265eca/download https://repository.ucc.edu.co/bitstreams/3293ddf8-91ee-4685-9c60-224d98ff5fb0/download https://repository.ucc.edu.co/bitstreams/5a1bee84-cb68-4d2a-8907-e84562a8f7fb/download https://repository.ucc.edu.co/bitstreams/23dad5f3-eaed-407f-b7c3-2ae393122b37/download |
| bitstream.checksum.fl_str_mv |
0e759f4ac494877ec8ebecde3a47dda3 1cb5e0ef6d42512b65e5a278e7e38be2 3bce4f7ab09dfc588f126e1e36e98a45 7aeab8de36990efab6b451de727e7c35 6c22d74eaa2b6d9d6f25f1423a20891c 99350a12e81c7c6a92f11e1021d2d087 237047f19095542365180903bc71ca39 |
| bitstream.checksumAlgorithm.fl_str_mv |
MD5 MD5 MD5 MD5 MD5 MD5 MD5 |
| repository.name.fl_str_mv |
Repositorio Institucional Universidad Cooperativa de Colombia |
| repository.mail.fl_str_mv |
bdigital@metabiblioteca.com |
| _version_ |
1811565417152380928 |
| spelling |
Vargas Bermúdez, DianaRendón Marín, SantiagoRuiz Sáenz, JuliánMogollón, JoséJaime, Jairo162022-02-15T20:20:49Z2022-02-15T20:20:49Z2021-121932-620310.1371/journal.pone.0258311https://hdl.handle.net/20.500.12494/43972Vargas-Bermudez DS, Rendon-Marin S, Ruiz-Saenz J, Mogollón D, Jaime J (2021) The first report of porcine parvovirus 7 (PPV7) in Colombia demonstrates the presence of variants associated with modifications at the level of the VP2-capsid protein. PLOS ONE 16(12): e0258311. https://doi.org/10.1371/journal.pone.0258311There are a wide variety of porcine parvoviruses (PPVs) referred to as PPV1 to PPV7. The latter was discovered in 2016 and later reported in some countries in America, Asia, and Europe. PPV7 as a pathogenic agent or coinfection with other pathogens causing disease has not yet been determined. In the present study, we report the identification of PPV7 for the first time in Colombia, where it was found retrospectively since 2015 in 40% of the provinces that make up the country (13/32), and the virus was ratified for 2018 in 4/5 provinces evaluated. Additionally, partial sequencing (nucleotides 380 to 4000) was performed of four Colombian strains completely covering the VP2 and NS1 viral genes. A sequence identity greater than 99% was found when comparing them with reference strains from the USA and China. In three of the four Colombian strains, an insertion of 15 nucleotides (five amino acids) was found in the PPV7-VP2 capsid protein (540–5554 nt; 180–184 aa). Based on this insertion, the VP2 phylogenetic analysis exhibited two well-differentiated evolutionarily related groups. To evaluate the impact of this insertion on the structure of the PPV7-VP2 capsid protein, the secondary structure of two different Colombian strains was predicted, and it was determined that the insertion is located in the coil region and not involved in significant changes in the structure of the protein. The 3D structure of the PPV7-VP2 capsid protein was determined by threading and homology modeling, and it was shown that the insertion did not imply a change in the shape of the protein. Additionally, it was determined that the insertion is not involved in suppressing a potential B cell epitope, although the increase in length of the epitope could affect the interaction with molecules that allow a specific immune response.http://scienti.colciencias.gov.co:8081/cvlac/visualizador/generarCurriculoCv.do?cod_rh=0000153095https://orcid.org/0000-0002-1447-1458https://scienti.minciencias.gov.co/gruplac/jsp/visualiza/visualizagr.jsp?nro=00000000000695julian.ruizs@campusucc.edu.cohttps://scholar.google.com/citations?user=o3Y7mZwAAAAJ&hl=ese0258311Universidad Cooperativa de Colombia, Facultad de Ciencias de la Salud, Medicina Veterinaría y Zootecnia, BucaramangaPLOSMedicina veterinaria y zootecniaBucaramangahttps://journals.plos.org/plosone/article?id=10.1371/journal.pone.0258311PLOS ONERanz AI, Manclús JJ, Díaz-Aroca E, Casal JI. Porcine parvovirus: DNA sequence and genome organization. J Gen Virol. 1989;70 (Pt 10): 2541–2553. pmid:2794971Pénzes JJ, Söderlund-Venermo M, Canuti M, Eis-Hübinger AM, Hughes J, Cotmore SF, et al. Reorganizing the family Parvoviridae: a revised taxonomy independent of the canonical approach based on host association. Arch Virol. 2020; pmid:32533329Yang S, Liu Z, Wang Y, Li W, Fu X, Lin Y, et al. A novel rodent Chapparvovirus in feces of wild rats. Virol J. 2016;13: 133. pmid:27473724Williams SH, Che X, Garcia JA, Klena JD, Lee B, Muller D, et al. Viral diversity of house mice in new york city. MBio. 2018;9. pmid:29666290Fahsbender E, Altan E, Seguin MA, Young P, Estrada M, Leutenegger C, et al. Chapparvovirus DNA Found in 4% of Dogs with Diarrhea. Viruses. 2019;11. pmid:31035625Reuter G, Boros Á, Delwart E, Pankovics P. Novel circular single-stranded DNA virus from turkey faeces. Arch Virol. 2014;159: 2161–2164. pmid:24562429Lima DA, Cibulski SP, Tochetto C, Varela APM, Finkler F, Teixeira TF, et al. The intestinal virome of malabsorption syndrome-affected and unaffected broilers through shotgun metagenomics. Virus Res. 2019;261: 9–20. pmid:30543873Souza WM de Romeiro MF, Fumagalli MJ, Modha S, de Araujo J, Queiroz LH, et al. Chapparvoviruses occur in at least three vertebrate classes and have a broad biogeographic distribution. J Gen Virol. 2017;98: 225–229. pmid:28284244Palinski RM, Mitra N, Hause BM. Discovery of a novel Parvovirinae virus, porcine parvovirus 7, by metagenomic sequencing of porcine rectal swabs. Virus Genes. 2016;52: 564–567. pmid:26995221Xing X, Zhou H, Tong L, Chen Y, Sun Y, Wang H, et al. First identification of porcine parvovirus 7 in China. Arch Virol. 2018;163: 209–213. pmid:29022179Wang Y, Yang K-K, Wang J, Wang X-P, Zhao L, Sun P, et al. Detection and molecular characterization of novel porcine parvovirus 7 in Anhui province from Central-Eastern China. Infect Genet Evol. 2019;71: 31–35. pmid:30876889Wang W, Cao L, Sun W, Xin J, Zheng M, Tian M, et al. Sequence and phylogenetic analysis of novel porcine parvovirus 7 isolates from pigs in Guangxi, China. PLoS One. 2019;14: e0219560. pmid:31291362Ouh I-O, Park S, Lee J-Y, Song JY, Cho I-S, Kim H-R, et al. First detection and genetic characterization of porcine parvovirus 7 from Korean domestic pig farms. J Vet Sci. 2018;19: 855–857. pmid:30304892Chung H-C, Nguyen V-G, Huynh T-M-L, Park Y-H, Park K-T, Park B-K. PCR-based detection and genetic characterization of porcine parvoviruses in South Korea in 2018. BMC Vet Res. 2020;16: 113. pmid:32295585Miłek D, Woźniak A, Stadejek T. The detection and genetic diversity of novel porcine parvovirus 7 (PPV7) on Polish pig farms. Res Vet Sci. 2018;120: 28–32. pmid:30170185Blomström A-L, Ye X, Fossum C, Wallgren P, Berg M. Characterisation of the Virome of Tonsils from Conventional Pigs and from Specific Pathogen-Free Pigs. Viruses. 2018;10. pmid:30036964Da Silva MS, Budaszewski RF, Weber MN, Cibulski SP, Paim WP, Mósena ACS, et al. Liver virome of healthy pigs reveals diverse small ssDNA viral genomes. Infect Genet Evol. 2020;81: 104203. pmid:32035977Shackelton LA, Parrish CR, Truyen U, Holmes EC. High rate of viral evolution associated with the emergence of carnivore parvovirus. Proc Natl Acad Sci USA. 2005;102: 379–384. pmid:15626758Streck AF, Canal CW, Truyen U. Molecular epidemiology and evolution of porcine parvoviruses. Infect Genet Evol. 2015;36: 300–306. pmid:26453771Ren X, Tao Y, Cui J, Suo S, Cong Y, Tijssen P. Phylogeny and evolution of porcine parvovirus. Virus Res. 2013;178: 392–397. pmid:24050995Wang D, Mai J, Yang Y, Wang N. Porcine Parvovirus 7: Evolutionary Dynamics and Identification of Epitopes toward Vaccine Design. Vaccines (Basel). 2020;8. pmid:32635618Streck AF, Bonatto SL, Homeier T, Souza CK, Gonçalves KR, Gava D, et al. High rate of viral evolution in the capsid protein of porcine parvovirus. J Gen Virol. 2011;92: 2628–2636. pmid:21795474Cadar D, Cságola A, Kiss T, Tuboly T. Capsid protein evolution and comparative phylogeny of novel porcine parvoviruses. Mol Phylogenet Evol. 2013;66: 243–253. pmid:23044400Cotmore SF, Agbandje-McKenna M, Chiorini JA, Mukha DV, Pintel DJ, Qiu J, et al. The family Parvoviridae. Arch Virol. 2014;159: 1239–1247. pmid:24212889Mengeling WL, Lager KM, Vorwald AC. The effect of porcine parvovirus and porcine reproductive and respiratory syndrome virus on porcine reproductive performance. Anim Reprod Sci. 2000;60–61: 199–210. pmid:10844195Miłek D, Woźniak A, Podgórska K, Stadejek T. Do porcine parvoviruses 1 through 7 (PPV1-PPV7) have an impact on porcine circovirus type 2 (PCV2) viremia in pigs? Vet Microbiol. 2020;242: 108613. pmid:32122579Opriessnig T, Fenaux M, Yu S, Evans RB, Cavanaugh D, Gallup JM, et al. Effect of porcine parvovirus vaccination on the development of PMWS in segregated early weaned pigs coinfected with type 2 porcine circovirus and porcine parvovirus. Vet Microbiol. 2004;98: 209–220. pmid:15036529Allan GM, Kennedy S, McNeilly F, Foster JC, Ellis JA, Krakowka SJ, et al. Experimental reproduction of severe wasting disease by co-infection of pigs with porcine circovirus and porcine parvovirus. J Comp Pathol. 1999;121: 1–11. pmid:10373289Ha Z, Xie C-Z, Li J-F, Wen S-B, Zhang K-L, Nan F-L, et al. Molecular detection and genomic characterization of porcine circovirus 3 in pigs from Northeast China. BMC Vet Res. 2018;14: 321. pmid:30367641Oraveerakul K, Choi CS, Molitor TW. Tissue tropisms of porcine parvovirus in swine. Arch Virol. 1993;130: 377–389. pmid:8390826Paul PS, Mengeling WL, Pirtle EC. Duration and biological half-life of passively acquired colostral antibodies to porcine parvovirus. Am J Vet Res. 1982;43: 1376–1379. pmid:7103222Agbandje-McKenna M, Llamas-Saiz AL, Wang F, Tattersall P, Rossmann MG. Functional implications of the structure of the murine parvovirus, minute virus of mice. Structure. 1998;6: 1369–1381. pmid:9817841Boisvert M, Fernandes S, Tijssen P. Multiple pathways involved in porcine parvovirus cellular entry and trafficking toward the nucleus. J Virol. 2010;84: 7782–7792. pmid:20484503Simpson AA, Hébert B, Sullivan GM, Parrish CR, Zádori Z, Tijssen P, et al. The structure of porcine parvovirus: comparison with related viruses. J Mol Biol. 2002;315: 1189–1198. pmid:11827486López-Bueno A, Rubio M-P, Bryant N, McKenna R, Agbandje-McKenna M, Almendral JM. Host-selected amino acid changes at the sialic acid binding pocket of the parvovirus capsid modulate cell binding affinity and determine virulence. J Virol. 2006;80: 1563–1573. pmid:16415031Ji P, Liu Y, Chen Y, Wang A, Jiang D, Zhao B, et al. Porcine parvovirus capsid protein expressed in Escherichia coli self-assembles into virus-like particles with high immunogenicity in mice and guinea pigs. Antiviral Res. 2017;139: 146–152. pmid:28063996Antonis AFG, Bruschke CJM, Rueda P, Maranga L, Casal JI, Vela C, et al. A novel recombinant virus-like particle vaccine for prevention of porcine parvovirus-induced reproductive failure. Vaccine. 2006;24: 5481–5490. pmid:16730104Hueffer K, Parker JSL, Weichert WS, Geisel RE, Sgro J-Y, Parrish CR. The natural host range shift and subsequent evolution of canine parvovirus resulted from virus-specific binding to the canine transferrin receptor. J Virol. 2003;77: 1718–1726. pmid:12525605Hu W, Xu X, Liu Q, Ji J, Kan Y, Yao L, et al. Molecular characterisation and genetic diversity of canine parvovirus type 2 prevalent in central china. J Vet Res. 2020;64: 347–354. pmid:32984622Gainor K, Bowen A, Bolfa P, Peda A, Malik YS, Ghosh S. Molecular Investigation of Canine Parvovirus-2 (CPV-2) Outbreak in Nevis Island: Analysis of the Nearly Complete Genomes of CPV-2 Strains from the Caribbean Region. Viruses. 2021;13. pmid:34204082Chen B, Zhang X, Zhu J, Liao L, Bao E. Molecular Epidemiological Survey of Canine Parvovirus Circulating in China from 2014 to 2019. Pathogens. 2021;10. pmid:34064982Parrish CR. Mapping specific functions in the capsid structure of canine parvovirus and feline panleukopenia virus using infectious plasmid clones. Virology. 1991;183: 195–205. pmid:1647068Kumar S, Stecher G, Tamura K. MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol Biol Evol. 2016;33: 1870–1874. pmid:27004904Jones DT. Protein secondary structure prediction based on position-specific scoring matrices. J Mol Biol. 1999;292: 195–202. pmid:10493868Roy A, Kucukural A, Zhang Y. I-TASSER: a unified platform for automated protein structure and function prediction. Nat Protoc. 2010;5: 725–738. pmid:20360767Yang J, Yan R, Roy A, Xu D, Poisson J, Zhang Y. The I-TASSER Suite: protein structure and function prediction. Nat Methods. 2015;12: 7–8. pmid:25549265Yang J, Zhang Y. I-TASSER server: new development for protein structure and function predictions. Nucleic Acids Res. 2015;43: W174–W181. pmid:25883148Eswar N, Webb B, Marti-Renom MA, Madhusudhan MS, Eramian D, Shen M-Y, et al. Comparative protein structure modeling using Modeller. Curr Protoc Bioinformatics. 2006;Chapter 5: Unit 5.6. pmid:18428767Xu D, Zhang Y. Improving the physical realism and structural accuracy of protein models by a two-step atomic-level energy minimization. Biophys J. 2011;101: 2525–2534. pmid:22098752Wiederstein M, Sippl MJ. ProSA-web: interactive web service for the recognition of errors in three-dimensional structures of proteins. Nucleic Acids Res. 2007;35: W407–10. pmid:17517781Sippl MJ. Recognition of errors in three-dimensional structures of proteins. Proteins. 1993;17: 355–362. pmid:8108378Laskowski RA, MacArthur MW, Moss DS, Thornton JM. PROCHECK: a program to check the stereochemical quality of protein structures. J Appl Crystallogr. 1993;26: 283–291.Zhang Y, Skolnick J. TM-align: a protein structure alignment algorithm based on the TM-score. Nucleic Acids Res. 2005;33: 2302–2309. pmid:15849316Bowie JU, Lüthy R, Eisenberg D. A method to identify protein sequences that fold into a known three-dimensional structure. Science. 1991;253: 164–170. pmid:1853201Lüthy R, Bowie JU, Eisenberg D. Assessment of protein models with three-dimensional profiles. Nature. 1992;356: 83–85. pmid:1538787Pettersen EF, Goddard TD, Huang CC, Couch GS, Greenblatt DM, Meng EC, et al. UCSF Chimera—a visualization system for exploratory research and analysis. J Comput Chem. 2004;25: 1605–1612. pmid:15264254Jespersen MC, Peters B, Nielsen M, Marcatili P. BepiPred-2.0: improving sequence-based B-cell epitope prediction using conformational epitopes. Nucleic Acids Res. 2017;45: W24–W29. pmid:28472356Yao B, Zhang L, Liang S, Zhang C. SVMTriP: a method to predict antigenic epitopes using support vector machine to integrate tri-peptide similarity and propensity. PLoS One. 2012;7: e45152. pmid:22984622CapsidsSwinePhylogenetic analysisB cellsNucleotide sequencingParvovirusesColombiaStructural proteinsThe first report of porcine parvovirus 7 (PPV7) in Colombia demonstrates the presence of variants associated with modifications at the level of the VP2-capsid proteinArtículos Científicoshttp://purl.org/coar/resource_type/c_2df8fbb1http://purl.org/coar/version/c_970fb48d4fbd8a85info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionAtribución – No comercial – Compartir igualinfo:eu-repo/semantics/openAccesshttp://purl.org/coar/access_right/c_abf2PublicationORIGINAL2022__first_report_porcine-FormatoLicenciaUso.pdf2022__first_report_porcine-FormatoLicenciaUso.pdfapplication/pdf207596https://repository.ucc.edu.co/bitstreams/c5a067e7-24ab-4ed6-8158-1ae42637e1dc/download0e759f4ac494877ec8ebecde3a47dda3MD512022__first_report_porcine.pdf2022__first_report_porcine.pdfapplication/pdf1599653https://repository.ucc.edu.co/bitstreams/8149ddcf-0f8b-4161-88d8-a01dfdc933e9/download1cb5e0ef6d42512b65e5a278e7e38be2MD52LICENSElicense.txtlicense.txttext/plain; charset=utf-84334https://repository.ucc.edu.co/bitstreams/8ca5238d-d9ba-4246-bae4-bc8ac9d3ea26/download3bce4f7ab09dfc588f126e1e36e98a45MD53THUMBNAIL2022__first_report_porcine-FormatoLicenciaUso.pdf.jpg2022__first_report_porcine-FormatoLicenciaUso.pdf.jpgGenerated Thumbnailimage/jpeg5422https://repository.ucc.edu.co/bitstreams/93819280-1a47-4f10-9c26-f0966f265eca/download7aeab8de36990efab6b451de727e7c35MD542022__first_report_porcine.pdf.jpg2022__first_report_porcine.pdf.jpgGenerated Thumbnailimage/jpeg5678https://repository.ucc.edu.co/bitstreams/3293ddf8-91ee-4685-9c60-224d98ff5fb0/download6c22d74eaa2b6d9d6f25f1423a20891cMD55TEXT2022__first_report_porcine-FormatoLicenciaUso.pdf.txt2022__first_report_porcine-FormatoLicenciaUso.pdf.txtExtracted texttext/plain6046https://repository.ucc.edu.co/bitstreams/5a1bee84-cb68-4d2a-8907-e84562a8f7fb/download99350a12e81c7c6a92f11e1021d2d087MD562022__first_report_porcine.pdf.txt2022__first_report_porcine.pdf.txtExtracted texttext/plain61980https://repository.ucc.edu.co/bitstreams/23dad5f3-eaed-407f-b7c3-2ae393122b37/download237047f19095542365180903bc71ca39MD5720.500.12494/43972oai:repository.ucc.edu.co:20.500.12494/439722024-08-09 12:27:19.771restrictedhttps://repository.ucc.edu.coRepositorio Institucional Universidad Cooperativa de Colombiabdigital@metabiblioteca.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 |