Whole genome characterization of Torque teno sus virus 1 (TTSuV1) in wild and domestic pigs: insights into genetic classification, host differentiation, and intra-host variation

BackgroundTorque teno sus virus 1 (TTSuV1), a member of the Anelloviridae family, is highly prevalent in swine populations and exhibits substantial genetic diversity. Despite its ubiquity, TTSuV1 remains understudied, particularly regarding its genetic diversity, host-specific differentiation, and i...

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Main Authors: Xiaolong Li, Yasmin Tavares, Céline M. Carneiro, Caroline Phillips, Kuttichantran Subramaniam, John Lednicky, Raoul K. Boughton, Kim M. Pepin, Ryan S. Miller, Kurt C. VerCauteren, Samantha M. Wisely
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Language:English
Published: Frontiers Media S.A. 2025-05-01
Series:Frontiers in Microbiology
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Online Access:https://www.frontiersin.org/articles/10.3389/fmicb.2025.1585558/full
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author Xiaolong Li
Yasmin Tavares
Céline M. Carneiro
Caroline Phillips
Kuttichantran Subramaniam
Kuttichantran Subramaniam
John Lednicky
John Lednicky
Raoul K. Boughton
Kim M. Pepin
Ryan S. Miller
Kurt C. VerCauteren
Samantha M. Wisely
Samantha M. Wisely
author_facet Xiaolong Li
Yasmin Tavares
Céline M. Carneiro
Caroline Phillips
Kuttichantran Subramaniam
Kuttichantran Subramaniam
John Lednicky
John Lednicky
Raoul K. Boughton
Kim M. Pepin
Ryan S. Miller
Kurt C. VerCauteren
Samantha M. Wisely
Samantha M. Wisely
author_sort Xiaolong Li
collection DOAJ
description BackgroundTorque teno sus virus 1 (TTSuV1), a member of the Anelloviridae family, is highly prevalent in swine populations and exhibits substantial genetic diversity. Despite its ubiquity, TTSuV1 remains understudied, particularly regarding its genetic diversity, host-specific differentiation, and intra-host variation. These characteristics are critical for understanding its evolution, transmission dynamics, and potential applications in biosecurity monitoring.MethodsField and laboratory protocols included capturing wild pigs, collecting whole blood samples, and screening for TTSuV1-positive samples through PCR. TOPO TA cloning was used to amplify individual viral variants within hosts, and whole genome sequencing (WGS) was performed on selected clones. A dated phylogenetic tree was reconstructed using TTSuV1 whole genome sequences obtained from wild pig samples in this study and all available sequences from NCBI. To evaluate genetic differentiation between wild and domestic pigs, partial viral sequences (~700 bp) were analyzed using phylogenetic D statistic and analysis of molecular variance (AMOVA). Intra-host variation was assessed by calculating pairwise identity percentages among viral clones from individual hosts and constructing haplotype networks.ResultsPhylogenetic analysis of whole genome sequences grouped TTSuV1 into four clades, with sequences from wild pigs distributed across all clades. Known subtypes 1a, 1b, and 1c were localized within Clades 3 and 4, leaving sequences in Clades 1 and 2 with unidentified subtypes. Partial sequence analysis revealed significant host-specific genetic differentiation: the D statistic confirmed a non-random association between host type (wild vs. domestic) and phylogeny, and AMOVA further showed contributions of both host type and geography to overall variation. Intra-host variation analysis provided evidence for multiple sources of genetic diversity within individual hosts. Pairwise identity percentages among viral clones ranged from 63.6% to 100%, with lower identity values indicating co-infection with distinct viral variants. Haplotype network analysis revealed mutational steps between haplotypes from the same host, suggesting that intra-host evolution also contributes to within-host genetic variation.ConclusionsThis study highlights the significant genetic diversity and host-specific differentiation of TTSuV1, with wild pigs playing a key role in its evolution. Both intra-host evolution and co-infection contribute to its diversity, underscoring its potential as a tool for monitoring biosecurity risks and cross-transmission between wild and domestic pigs.
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spelling doaj-art-55f3d31eaae344879b4d1d8bc73186ec2025-08-20T02:28:20ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2025-05-011610.3389/fmicb.2025.15855581585558Whole genome characterization of Torque teno sus virus 1 (TTSuV1) in wild and domestic pigs: insights into genetic classification, host differentiation, and intra-host variationXiaolong Li0Yasmin Tavares1Céline M. Carneiro2Caroline Phillips3Kuttichantran Subramaniam4Kuttichantran Subramaniam5John Lednicky6John Lednicky7Raoul K. Boughton8Kim M. Pepin9Ryan S. Miller10Kurt C. VerCauteren11Samantha M. Wisely12Samantha M. Wisely13Department of Wildlife Ecology and Conservation, University of Florida, Gainesville, FL, United StatesDepartment of Wildlife Ecology and Conservation, University of Florida, Gainesville, FL, United StatesDepartment of Wildlife Ecology and Conservation, University of Florida, Gainesville, FL, United StatesDepartment of Wildlife Ecology and Conservation, University of Florida, Gainesville, FL, United StatesDepartment of Infectious Diseases and Immunology, College of Veterinary Medicine, University of Florida, Gainesville, FL, United StatesEmerging Pathogens Institute, University of Florida, Gainesville, FL, United StatesEmerging Pathogens Institute, University of Florida, Gainesville, FL, United StatesDepartment of Environmental and Global Health, College of Public Health and Health Professions, University of Florida, Gainesville, FL, United StatesBuck Island Ranch, Archbold Biological Station, Lake Placid, FL, United StatesNational Wildlife Research Center, United States Department of Agriculture, Animal and Plant Health Inspection Service, Wildlife Services, Fort Collins, CO, United StatesCenter for Epidemiology and Animal Health, United States Department of Agriculture, Animal and Plant Health Inspection Service, Veterinary Services, Fort Collins, CO, United StatesNational Wildlife Research Center, United States Department of Agriculture, Animal and Plant Health Inspection Service, Wildlife Services, Fort Collins, CO, United StatesDepartment of Wildlife Ecology and Conservation, University of Florida, Gainesville, FL, United StatesEmerging Pathogens Institute, University of Florida, Gainesville, FL, United StatesBackgroundTorque teno sus virus 1 (TTSuV1), a member of the Anelloviridae family, is highly prevalent in swine populations and exhibits substantial genetic diversity. Despite its ubiquity, TTSuV1 remains understudied, particularly regarding its genetic diversity, host-specific differentiation, and intra-host variation. These characteristics are critical for understanding its evolution, transmission dynamics, and potential applications in biosecurity monitoring.MethodsField and laboratory protocols included capturing wild pigs, collecting whole blood samples, and screening for TTSuV1-positive samples through PCR. TOPO TA cloning was used to amplify individual viral variants within hosts, and whole genome sequencing (WGS) was performed on selected clones. A dated phylogenetic tree was reconstructed using TTSuV1 whole genome sequences obtained from wild pig samples in this study and all available sequences from NCBI. To evaluate genetic differentiation between wild and domestic pigs, partial viral sequences (~700 bp) were analyzed using phylogenetic D statistic and analysis of molecular variance (AMOVA). Intra-host variation was assessed by calculating pairwise identity percentages among viral clones from individual hosts and constructing haplotype networks.ResultsPhylogenetic analysis of whole genome sequences grouped TTSuV1 into four clades, with sequences from wild pigs distributed across all clades. Known subtypes 1a, 1b, and 1c were localized within Clades 3 and 4, leaving sequences in Clades 1 and 2 with unidentified subtypes. Partial sequence analysis revealed significant host-specific genetic differentiation: the D statistic confirmed a non-random association between host type (wild vs. domestic) and phylogeny, and AMOVA further showed contributions of both host type and geography to overall variation. Intra-host variation analysis provided evidence for multiple sources of genetic diversity within individual hosts. Pairwise identity percentages among viral clones ranged from 63.6% to 100%, with lower identity values indicating co-infection with distinct viral variants. Haplotype network analysis revealed mutational steps between haplotypes from the same host, suggesting that intra-host evolution also contributes to within-host genetic variation.ConclusionsThis study highlights the significant genetic diversity and host-specific differentiation of TTSuV1, with wild pigs playing a key role in its evolution. Both intra-host evolution and co-infection contribute to its diversity, underscoring its potential as a tool for monitoring biosecurity risks and cross-transmission between wild and domestic pigs.https://www.frontiersin.org/articles/10.3389/fmicb.2025.1585558/fullTorque teno sus virus 1wild pigsgenetic diversityhost-specific differentiationintra-host variation
spellingShingle Xiaolong Li
Yasmin Tavares
Céline M. Carneiro
Caroline Phillips
Kuttichantran Subramaniam
Kuttichantran Subramaniam
John Lednicky
John Lednicky
Raoul K. Boughton
Kim M. Pepin
Ryan S. Miller
Kurt C. VerCauteren
Samantha M. Wisely
Samantha M. Wisely
Whole genome characterization of Torque teno sus virus 1 (TTSuV1) in wild and domestic pigs: insights into genetic classification, host differentiation, and intra-host variation
Frontiers in Microbiology
Torque teno sus virus 1
wild pigs
genetic diversity
host-specific differentiation
intra-host variation
title Whole genome characterization of Torque teno sus virus 1 (TTSuV1) in wild and domestic pigs: insights into genetic classification, host differentiation, and intra-host variation
title_full Whole genome characterization of Torque teno sus virus 1 (TTSuV1) in wild and domestic pigs: insights into genetic classification, host differentiation, and intra-host variation
title_fullStr Whole genome characterization of Torque teno sus virus 1 (TTSuV1) in wild and domestic pigs: insights into genetic classification, host differentiation, and intra-host variation
title_full_unstemmed Whole genome characterization of Torque teno sus virus 1 (TTSuV1) in wild and domestic pigs: insights into genetic classification, host differentiation, and intra-host variation
title_short Whole genome characterization of Torque teno sus virus 1 (TTSuV1) in wild and domestic pigs: insights into genetic classification, host differentiation, and intra-host variation
title_sort whole genome characterization of torque teno sus virus 1 ttsuv1 in wild and domestic pigs insights into genetic classification host differentiation and intra host variation
topic Torque teno sus virus 1
wild pigs
genetic diversity
host-specific differentiation
intra-host variation
url https://www.frontiersin.org/articles/10.3389/fmicb.2025.1585558/full
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