Virome analysis of three Ixodidae ticks species from Colombia: A potential strategy for discovering and surveying tick-borne viruses
Ticks are a group of obligate blood-sucking ectoparasites that play a critical role in transmitting several important zoonotic pathogens that can infect animals and humans. Viruses are part of the tick microbiome and are involved in the transmission of important diseases. Furthermore, the little inf...
- Autores:
-
Orozco Orozco, Mateo
Gómez García, Giovan Fernando
Alzate Restrepo, Juan Fernando
Isaza Agudelo, Juan Pablo
Gutiérrez Builes, Lina Andrea
- Tipo de recurso:
- Article of investigation
- Fecha de publicación:
- 2021
- Institución:
- Tecnológico de Antioquia
- Repositorio:
- Repositorio Tdea
- Idioma:
- eng
- OAI Identifier:
- oai:dspace.tdea.edu.co:tdea/2771
- Acceso en línea:
- https://dspace.tdea.edu.co/handle/tdea/2771
- Palabra clave:
- Rhipicephalus
Rhipicephalus sanguineus
Garrapata Marrón del Perro
Zoonosis
Tick-borne diseases
Enfermedades por Picaduras de Garrapatas
Doenças Transmitidas por Carrapatos
Rhipicephalus microplus
Dermacentor nitens
- Rights
- openAccess
- License
- https://creativecommons.org/licenses/by-nc-nd/4.0/
id |
RepoTdea2_e39bc68f21968fd6cc95f6fb21af6b93 |
---|---|
oai_identifier_str |
oai:dspace.tdea.edu.co:tdea/2771 |
network_acronym_str |
RepoTdea2 |
network_name_str |
Repositorio Tdea |
repository_id_str |
|
dc.title.none.fl_str_mv |
Virome analysis of three Ixodidae ticks species from Colombia: A potential strategy for discovering and surveying tick-borne viruses |
title |
Virome analysis of three Ixodidae ticks species from Colombia: A potential strategy for discovering and surveying tick-borne viruses |
spellingShingle |
Virome analysis of three Ixodidae ticks species from Colombia: A potential strategy for discovering and surveying tick-borne viruses Rhipicephalus Rhipicephalus sanguineus Garrapata Marrón del Perro Zoonosis Tick-borne diseases Enfermedades por Picaduras de Garrapatas Doenças Transmitidas por Carrapatos Rhipicephalus microplus Dermacentor nitens |
title_short |
Virome analysis of three Ixodidae ticks species from Colombia: A potential strategy for discovering and surveying tick-borne viruses |
title_full |
Virome analysis of three Ixodidae ticks species from Colombia: A potential strategy for discovering and surveying tick-borne viruses |
title_fullStr |
Virome analysis of three Ixodidae ticks species from Colombia: A potential strategy for discovering and surveying tick-borne viruses |
title_full_unstemmed |
Virome analysis of three Ixodidae ticks species from Colombia: A potential strategy for discovering and surveying tick-borne viruses |
title_sort |
Virome analysis of three Ixodidae ticks species from Colombia: A potential strategy for discovering and surveying tick-borne viruses |
dc.creator.fl_str_mv |
Orozco Orozco, Mateo Gómez García, Giovan Fernando Alzate Restrepo, Juan Fernando Isaza Agudelo, Juan Pablo Gutiérrez Builes, Lina Andrea |
dc.contributor.author.none.fl_str_mv |
Orozco Orozco, Mateo Gómez García, Giovan Fernando Alzate Restrepo, Juan Fernando Isaza Agudelo, Juan Pablo Gutiérrez Builes, Lina Andrea |
dc.subject.decs.none.fl_str_mv |
Rhipicephalus Rhipicephalus sanguineus Garrapata Marrón del Perro Zoonosis Tick-borne diseases Enfermedades por Picaduras de Garrapatas Doenças Transmitidas por Carrapatos |
topic |
Rhipicephalus Rhipicephalus sanguineus Garrapata Marrón del Perro Zoonosis Tick-borne diseases Enfermedades por Picaduras de Garrapatas Doenças Transmitidas por Carrapatos Rhipicephalus microplus Dermacentor nitens |
dc.subject.proposal.none.fl_str_mv |
Rhipicephalus microplus Dermacentor nitens |
description |
Ticks are a group of obligate blood-sucking ectoparasites that play a critical role in transmitting several important zoonotic pathogens that can infect animals and humans. Viruses are part of the tick microbiome and are involved in the transmission of important diseases. Furthermore, the little information on these as etiological agents of zoonoses suggests the need to study these microorganisms. For this reason, in this study, we sought to characterize the virome in Rhipicephalus microplus, Dermacentor nitens, and Rhipicephalus sanguineus s.l., which were collected from different domestic animals in Antioquia, Colombia. RNA sequencing was used for virome characterization in these three tick species, using RNA-dependent polymerase as a marker gene. Forty-eight sequences corresponding to 14 different viruses were identified, some of which were previously identified in the tick’s virome. Overall, these data indicate that ticks from domestic animals in cattle farms harbor a wide viral diversity at the local scale. Thus, the metatranscriptomic approach provides important baseline information for monitoring the tick virome and to develop future studies on their biology, host-virus interactions, host range, worldwide distribution, and finally, their potential role as emerging vector-borne agents |
publishDate |
2021 |
dc.date.issued.none.fl_str_mv |
2021 |
dc.date.accessioned.none.fl_str_mv |
2023-04-11T19:16:12Z |
dc.date.available.none.fl_str_mv |
2023-04-11T19:16:12Z |
dc.type.spa.fl_str_mv |
Artículo de revista |
dc.type.coar.spa.fl_str_mv |
http://purl.org/coar/resource_type/c_2df8fbb1 |
dc.type.content.spa.fl_str_mv |
Text |
dc.type.driver.spa.fl_str_mv |
info:eu-repo/semantics/article |
dc.type.redcol.spa.fl_str_mv |
http://purl.org/redcol/resource_type/ART |
dc.type.version.spa.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.coarversion.spa.fl_str_mv |
http://purl.org/coar/version/c_970fb48d4fbd8a85 |
format |
http://purl.org/coar/resource_type/c_2df8fbb1 |
status_str |
publishedVersion |
dc.identifier.issn.spa.fl_str_mv |
1567-1348 |
dc.identifier.uri.none.fl_str_mv |
https://dspace.tdea.edu.co/handle/tdea/2771 |
dc.identifier.eissn.spa.fl_str_mv |
1567-7257 |
identifier_str_mv |
1567-1348 1567-7257 |
url |
https://dspace.tdea.edu.co/handle/tdea/2771 |
dc.language.iso.spa.fl_str_mv |
eng |
language |
eng |
dc.relation.citationendpage.spa.fl_str_mv |
10 |
dc.relation.citationstartpage.spa.fl_str_mv |
1 |
dc.relation.citationvolume.spa.fl_str_mv |
96 |
dc.relation.ispartofjournal.spa.fl_str_mv |
Infection, genetics and evolution |
dc.relation.references.spa.fl_str_mv |
Adams, M., Kreuze, J., Martelli, G., 2011. Tymovirales [WWW Document]. Int. Comm. Taxon. Viruses ICTV Altschul, S.F., Gish, W., Miller, W., Myers, E.W., Lipman, D.J., 1990. Basic local alignment search tool. J. Mol. Biol. 215, 403–410. https://doi.org/10.1016/S0022- 2836(05)80360-2. Alvarez, V., Bonilla, R., Chac ́on, I., 2003. Frecuencia relativa de Boophilus microplus (Acari: Ixodidae) en bovinos (Bos taurus y B. indicus) en ocho zonas ecol ́ogicas de Costa Rica. Rev. Biol. Trop. 51, 427–434. Andrews, S., 2010. FastQC: A Quality Control Tool for High Throughput Sequence Data [WWW Document]. Online. URL. http://www.bioinformatics.babraham.ac.uk/pro jects/fastqc/. Betancur, O.H., Betancourt, A.E., Giraldo, C.R., 2015. Importance of ticks in the transmission of zoonotic agents Importancia de las garrapatas en la transmisi ́on de agentes causantes de zoonosis. Rev. MVZ C ́ordoba 20, 5053–5067. Bolger, A.M., Lohse, M., Usadel, B., 2014. Trimmomatic: a flexible trimmer for Illumina sequence data. Bioinformatics 30, 2114–2120. https://doi.org/10.1093/ bioinformatics/btu170. Bonnet, S.I., Pollet, T., 2021. Update on the intricate tango between tick microbiomes and tick-borne pathogens. Parasite Immunol. 43, e12813 https://doi.org/10.1111/ pim.12813. Brinkmann, A., Nitsche, A., Kohl, C., 2016. Viral metagenomics on blood-feeding arthropods as a tool for human disease surveillance. Int. J. Mol. Sci. 17, E1743 https://doi.org/10.3390/ijms17101743. Buchfink, B., Xie, C., Huson, D.H., 2015. Fast and sensitive protein alignment using DIAMOND. Nat. Methods 12, 59–60. https://doi.org/10.1038/nmeth.3176. C ́amara de Comercio de Medellín para Antioquia, 2017. Perfil Socioecon ́omico de la Subregi ́on Magdalena Medio [WWW Document]. Inf. Estud. Econ ́omicos. URL https ://www.camaramedellin.com.co/Portals/0/Biblioteca/Estudios-economicos/cade nas-productivas-regionales/12-3Perfil Magdalena_Oct14.pdf?ver=2019-03-01- 095037-233 (accessed 3.3.21). Colombo, V.C., Fasano, A.A., Beldomenico, P.M., Nava, S., 2018. Tick host specificity: an analysis based on host phylogeny and tick ecological features using Amblyomma triste and Amblyomma tigrinum immature stages. Ticks Tick. Borne. Dis. 9, 781–787. https://doi.org/10.1016/j.ttbdis.2018.03.001. Cross, S.T., Kapuscinski, M.L., Perino, J., Maertens, B.L., Weger-Lucarelli, J., Ebel, G.D., Stenglein, M.D., 2018. Co-infection patterns in individual Ixodes scapularis ticks reveal associations between viral, eukaryotic and bacterial microorganisms. Viruses 10, 388. https://doi.org/10.3390/v10070388. de Souza, W.M., Fumagalli, M.J., Carrasco, Torres, Romeiro, M.F., Modha, S., Seki, M.C., Gheller, J.M., Daffre, S., Nunes, M.R.T., Murcia, P.R., Acrani, G.O., Figueiredo, L.T. M., 2018. Viral diversity of Rhipicephalus microplus parasitizing cattle in southern Brazil. Sci. Rep. 8, 16315. https://doi.org/10.1038/s41598-018-34630-1. Easton, A.J., Pringle, C.R., 2011. Mononegavirales [WWW Document]. Noveno Inf. ICTV. Eddy, S.R., 2011. Accelerated profile HMM searches. PLoS Comput. Biol. 7. https://doi. org/10.1371/journal.pcbi.1002195. Edgar, R.C., 2004. MUSCLE: multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Res. 32, 1792–1797. https://doi.org/10.1093/nar/ gkh340. Esser, H.J., Herre, E.A., Kays, R., Liefting, Y., Jansen, P.A., 2019. Local host-tick coextinction in neotropical forest fragments. Int. J. Parasitol. 49, 225–233. https:// doi.org/10.1016/j.ijpara.2018.08.008. Estrada-Pe ̃na, A., 2008. Climate, niche, ticks, and models: what they are and how we should interpret them. Parasitol. Res. 103, S87–S95. https://doi.org/10.1007/ s00436-008-1056-7 Estrada-Pe ̃na, A., Nava, S., Tarragona, E., de la Fuente, J., Guglielmone, A.A., 2020. A community approach to the Neotropical ticks-hosts interactions. Sci. Rep. 10, 9269. https://doi.org/10.1038/s41598-020-66400-3. Faizah, A.N., Kobayashi, D., Isawa, H., Amoa-Bosompem, M., Murota, K., Higa, Y., Futami, K., Shimada, S., Kim, K.S., Itokawa, K., Watanabe, M., Tsuda, Y., Minakawa, N., Miura, K., Hirayama, K., Sawabe, K., 2020. Deciphering the virome of Culex vishnui subgroup mosquitoes, the major vectors of japanese encephalitis, in Japan. Viruses 12, 264. https://doi.org/10.3390/v12030264. G ́omez, G.F., Isaza, J.P., Segura, J.A., Alzate, J.F., Guti ́errez, L.A., 2020. Metatranscriptomic virome assessment of Rhipicephalus microplus from Colombia. Ticks Tick. Borne. Dis. 11, 101426. https://doi.org/10.1016/j.ttbdis.2020.101426. Gondard, M., Temmam, S., Devillers, E., Bigot, T., Chr, D., Aprelon, R., Vayssier- taussat, M., Albina, E., Eloit, M., Moutailler, S., 2020. RNA Viruses of Amblyomma Variegatum and Rhipicephalus Microplus and Cattle Susceptibility in the French Antilles, pp. 1–23 Grabherr, M.G., Haas, B.J., Yassour, M., Levin, J.Z., Thompson, D.A., Amit, I., Adiconis, X., Fan, L., Raychowdhury, R., Zeng, Q., Chen, Z., Mauceli, E., Hacohen, N., Gnirke, A., Rhind, N., di Palma, F., Birren, B.W., Nusbaum, C., Lindblad Toh, K., Friedman, N., Regev, A., 2013. Trinity: reconstructing a full-length transcriptome without a genome from RNA-Seq data. Nat. Biotechnol. 29, 644–652. https://doi.org/10.1038/nbt.1883.Trinity Grard, G., Moureau, G., Charrel, R.N., Lemasson, J.J., Gonzalez, J.P., Gallian, P., Gritsun, T.S., Holmes, E.C., Gould, E.A., de Lamballerie, X., 2007. Genetic characterization of tick-borne flaviviruses: new insights into evolution, pathogenetic determinants and taxonomy. Virology 361, 80–92. https://doi.org/10.1016/j. virol.2006.09.015. Guglielmone, A.A., Apanaskevich, D.A., Estrada-Pe ̃na, A., Robbins, R.G., Petney, T.N., Horak, I.G., 2014. The hard ticks of the world: (Acari: Ixodida: Ixodidae). In: The Hard Ticks of the World: (Acari: Ixodida: Ixodidae). Springer Netherlands, Dordrecht, The Netherlands. https://doi.org/10.1007/978-94-007-7497-1. Harris, H.M.B., Hill, C., 2021. A place for viruses on the tree of life. Front. Microbiol. 11, 604048. https://doi.org/10.3389/fmicb.2020.604048. Huson, D.H., Auch, A.F., Qi, J., Schuster, S.C., 2007. MEGAN analysis of metagenomic data. Genome Res. 17, 377–386. https://doi.org/10.1101/gr.5969107. Hyatt, D., Locascio, P.F., Hauser, L.J., Uberbacher, E.C., 2012. Gene and translation initiation site prediction in metagenomic sequences. Bioinformatics 28, 2223–2230. https://doi.org/10.1093/bioinformatics/bts429. International Committee on Taxonomy of Viruses, 2012. Tymoviridae. In: King, A.M.Q., Adams, M.J., Carstens, E.B., Lefkowitz, E.J. (Eds.), Virus Taxonomy. Elsevier, pp. 944–952. https://doi.org/10.1016/b978-0-12-384684-6.00080-x. Kazimírov ́a, M., Thangamani, S., Bartíkov ́a, P., Hermance, M., Holíkov ́a, V., ˇStibr ́aniov ́a, I., Nuttall, P.A., 2017. Tick-borne viruses and biological processes at the tick-host-virus interface. Front. Cell. Infect. Microbiol. 7, 1–21. https://doi.org/ 10.3389/fcimb.2017.00339. Kazimírov ́a, M., Bartíkov ́a, P., ˇStibr ́aniov ́a, I., 2018. Tick-borne viruses and host skin interface. In: Skin and Arthropod Vectors. Elsevier Inc, pp. 325–383. https://doi.org/ 10.1016/B978-0-12-811436-0.00010-1. Kumar, A., Murthy, S., Kapoor, A., 2017. Evolution of selective-sequencing approaches for virus discovery and virome analysis. Virus Res. https://doi.org/10.1016/j. virusres.2017.06.005. Letunic, I., Bork, P., 2021. Interactive tree of life (iTOL) v5: an online tool for phylogenetic tree display and annotation. Nucleic Acids Res. https://doi.org/ 10.1093/nar/gkab301 gkab301 Li, H., Durbin, R., 2009. Fast and accurate short read alignment with burrows-wheeler transform. Bioinformatics 25, 1754–1760. https://doi.org/10.1093/bioinformatics/ btp324. Li, C.X., Shi, M., Tian, J.H., Lin, X.D., Kang, Y.J., Chen, L.J., Qin, X.C., Xu, J., Holmes, E. C., Zhang, Y.Z., 2015. Unprecedented genomic diversity of RNA viruses in arthropods reveals the ancestry of negative-sense RNA viruses. Elife 4, e05378. https://doi.org/10.7554/eLife.05378. Liu, Q., He, B., Huang, S.Y., Wei, F., Zhu, X.Q., 2014. Severe fever with thrombocytopenia syndrome, an emerging tick-borne zoonosis. Lancet Infect. Dis. 14, 763–772. https://doi.org/10.1016/S1473-3099(14)70718-2. L ́opez, Y., Miranda, J., Mattar, S., Gonzalez, M., Rovnak, J., 2020. First report of Lihan Tick virus (Phlebovirus, Phenuiviridae) in ticks, Colombia. Virol. J. 17, 1–6. https:// doi.org/10.1186/s12985-020-01327-9. Madison-Antenucci, S., Kramer, L.D., Gebhardt, L.L., Kauffman, E., 2020. Emerging tick- borne diseases. Clin. Microbiol. Rev. https://doi.org/10.1128/CMR e00083–18. Martelli, G.P., Adams, M.J., Kreuze, J.F., Dolja, V.V., 2008. Family Flexiviridae: a case study in virion and genome plasticity. Annu. Rev. Phytopathol. 45, 73–100. https:// doi.org/10.1146/annurev.phyto.45.062806.094401. Meng, F., Ding, M., Tan, Z., Zhao, Z., Xu, L., Wu, J., He, B., Tu, C., 2018. Virome analysis of tick-borne viruses in Heilongjiang Province, China. Ticks Tick. Borne. Dis. 10, 412–420. https://doi.org/10.1016/j.ttbdis.2018.12.002. Monsalve-Arteagaid, L., Alonso-Sard ́on, M., Bellido, J.L.M., Santiago, M.B.V., Lista, M.C. V., Ab ́an, J.L., Muro, A., Belhassen-García, M., 2020. Seroprevalence of crimean- congo hemorrhagic fever in humans in the world health organization european region: a systematic review. PLoS Negl. Trop. Dis. 14, e0008094 https://doi.org/ 10.1371/journal.pntd.0008094. Morrone, J.J., 2014. Biogeographical regionalisation of the Neotropical region. Zootaxa 3782, 1–110. https://doi.org/10.11646/zootaxa.3782.1.1. Narasimhan, S., Fikrig, E., 2015. Tick microbiome: the force within. Trends Parasitol. 31, 315–323. https://doi.org/10.1016/j.pt.2015.03.010. Nava, S., Guglielmone, A.A., 2013. A meta-analysis of host specificity in Neotropical hard ticks (Acari: Ixodidae). Bull. Entomol. Res. 103, 216–224. https://doi.org/10.1017/ S0007485312000557 Ortíz-Giraldo, M., Tob ́on-Escobar, W.D., Vel ́asquez-Guarín, D., Usma-Marín, M.F., Ossa- L ́opez, P.A., Ramírez-Chaves, H.E., Carvajal-Agudelo, J.D., Rivera-P ́aez, F.A., 2021. Ticks (Acari: Ixodoidea) associated with mammals in Colombia: a historical review, molecular species confirmation, and establishment of new relationships. Parasitol. Res. 120, 383–394. https://doi.org/10.1007/s00436-020-06989-6. Paules, C.I., Marston, H.D., Bloom, M.E., Fauci, A.S., 2018. Tickborne diseases — confronting a growing threat. N. Engl. J. Med. 379, 701–703. https://doi.org/ 10.1056/nejmp1807870. Prasad, N., Murdoch, D.R., Reyburn, H., Crump, J.A., 2015. Etiology of severe febrile illness in low- and middle-income countries: a systematic review. PLoS One 10, e0127962. https://doi.org/10.1371/journal.pone.0127962 Regier, Y., Komma, K., Weigel, M., Kraiczy, P., Laisi, A., Pulliainen, A.T., Hain, T., Kempf, V.A.J., 2019. Combination of microbiome analysis and serodiagnostics to assess the risk of pathogen transmission by ticks to humans and animals in Central Germany 11 medical and health sciences 1108 medical microbiology. Parasit. Vectors 12. https://doi.org/10.1186/s13071-018-3240-7 Rosenberg, R., Lindsey, N.P., Fischer, M., Gregory, C.J., Hinckley, A.F., Mead, P.S., Paz- Bailey, G., Waterman, S.H., Drexler, N.A., Kersh, G.J., Hooks, H., Partridge, S.K., Visser, S.N., Beard, C.B., Petersen, L.R., 2018. Vital signs: trends in reported vectorborne disease cases — United States and territories, 2004–2016. MMWR Morb. Mortal. Wkly Rep. 67, 496–501. https://doi.org/10.15585/mmwr. mm6717e1. Sameroff, S., Tokarz, R., Charles, R.A., Jain, K., Oleynik, A., Che, X., Georges, K., Carrington, C.V., Lipkin, W.I., Oura, C., 2019. Viral diversity of tick species parasitizing cattle and dogs in Trinidad and Tobago. Sci. Rep. 9, 10421. https://doi. org/10.1038/s41598-019-46914-1. Savage, H.M., Godsey, M.S., Amy, L., Panella, N.A., Burkhalter, K.L., Harmon, J.R., Lash, R.R., Ashley, D.C., Nicholson, W.L., 2013. First detection of Heartland virus (bunyaviridae: Phlebovirus) from field collected arthropods. Am. J. Trop. Med. Hyg. 89, 445–452. https://doi.org/10.4269/ajtmh.13-0209. . Segura, J.A., Isaza, J.P., Botero, L.E., Alzate, J.F., Guti ́errez, L.A., 2020. Assessment of bacterial diversity of Rhipicephalus microplus ticks from two livestock agroecosystems in Antioquia, Colombia. PLoS One 15. https://doi.org/10.1371/ journal.pone.0234005. Shi, M., Lin, X.D., Tian, J.H., Chen, L.J., Chen, X., Li, C.X., Qin, X.C., Li, J., Cao, J.P., Eden, J.S., Buchmann, J., Wang, W., Xu, J., Holmes, E.C., Zhang, Y.Z., 2016. Redefining the invertebrate RNA virosphere. Nature 540, 539–543. https://doi.org/ 10.1038/nature20167 Shi, J., Hu, Z., Deng, F., Shen, S., 2018. Tick-borne viruses. Virol. Sin. 33, 21–43. https:// doi.org/10.1007/s12250-018-0019-0. Shi, J., Shen, S., Wu, H., Zhang, Y., Deng, F., 2021. Metagenomic profiling of viruses associated with Rhipicephalus microplus ticks in Yunnan Province, China. Virol. Sin. https://doi.org/10.1007/s12250-020-00319-x. Stamatakis, A., 2014. RAxML version 8: a tool for phylogenetic analysis and post-analysis of large phylogenies. Bioinformatics 30, 1312–1313. https://doi.org/10.1093/ bioinformatics/btu033. Segura, J.A., Isaza, J.P., Botero, L.E., Alzate, J.F., Guti ́errez, L.A., 2020. Assessment of bacterial diversity of Rhipicephalus microplus ticks from two livestock agroecosystems in Antioquia, Colombia. PLoS One 15. https://doi.org/10.1371/ journal.pone.0234005 Starr, E.P., Nuccio, E.E., Pett-Ridge, J., Banfield, J.F., Firestone, M.K., 2019. Metatranscriptomic reconstruction reveals RNA viruses with the potential to shape carbon cycling in soil. bioRxiv 116, 25900–25908. https://doi.org/10.1101/ 597468. Temmam, S., Chr ́etien, D., Bigot, T., Dufour, E., Petres, S., Desquesnes, M., Devillers, E., Dumarest, M., Yousfi, L., Jittapalapong, S., Karnchanabanthoeng, A., Chaisiri, K., Gagnieur, L., Cosson, J.F., Vayssier-Taussat, M., Morand, S., Moutailler, S., Eloit, M., 2019. Monitoring silent spillovers before emergence: a pilot study at the tick/human interface in Thailand. Front. Microbiol. 10, 2315. https://doi.org/10.3389/ fmicb.2019.02315. Tokarz, R., Williams, S.H., Sameroff, S., Sanchez Leon, M., Jain, K., Lipkin, W.I., 2014. Virome analysis of Amblyomma americanum, Dermacentor variabilis, and Ixodes scapularis ticks reveals novel highly divergent vertebrate and invertebrate viruses. J. Virol. 88, 11480–11492. https://doi.org/10.1128/JVI.01858-14. Vandegrift, K.J., Kapoor, A., 2019. The ecology of new constituents of the tick virome and their relevance to public health. Viruses 11, 529. https://doi.org/10.3390/ v11060529. Walker, A.R., Bouattour, A., Camicas, J., Estrada-Pe ̃na, A., Horak, I.G., Latif, A.A., Pegram, R.G., Preston, P.M., 2003. Ticks of Domestic Animals in Africa: A Guide to Identification of Species. Bioscience Reports, Edinburgh Scotland,U.K. Wang, L., Lv, X., Zhai, Y., Fu, S., Wang, D., Rayner, S., Tang, Q., Liang, G., 2012. Genomic characterization of a novel virus of the family Tymoviridae isolated from mosquitoes. PLoS One 7. https://doi.org/10.1371/journal.pone.0039845. Waterhouse, R.M., Seppey, M., Simao, F.A., Manni, M., Ioannidis, P., Klioutchnikov, G., Kriventseva, E.V., Zdobnov, E., 2018. BUSCO applications from quality assessments to gene prediction and phylogenomics. Mol. Biol. Evol. 35, 543–548. https://doi. org/10.1093/molbev/msx319 Wolf, Y.I., Kazlauskas, D., Iranzo, J., Luc ́Ia-Sanz, A., Kuhn, J.H., Krupovic, M., Dolja, V. V., Koonin, E.V., 2018. Origins and evolution of the global RNA virome. MBio 9, 1–31. https://doi.org/10.1101/451740 . Wu, H., Pang, R., Cheng, T., Xue, L., Zeng, H., Lei, T., Chen, M., Wu, S., Ding, Y., Zhang, J., Shi, M., Wu, Q., 2020. Abundant and diverse RNA viruses in insects revealed by RNA-Seq analysis: ecological and evolutionary implications. mSystems 5. https://doi.org/10.1128/msystems.00039-20. Zhao, T., Gong, H., Shen, X., Zhang, W., Shan, T., Yu, X., Wang, S.J., Cui, L., 2020. Comparison of viromes in ticks from fifferent domestic animals in China. Virol. Sin. 35, 398–406. https://doi.org/10.1007/s12250-020-00197-3. |
dc.rights.uri.spa.fl_str_mv |
https://creativecommons.org/licenses/by-nc-nd/4.0/ |
dc.rights.license.spa.fl_str_mv |
Atribución-NoComercial-SinDerivadas 4.0 Internacional (CC BY-NC-ND 4.0) |
dc.rights.accessrights.spa.fl_str_mv |
info:eu-repo/semantics/openAccess |
dc.rights.coar.spa.fl_str_mv |
http://purl.org/coar/access_right/c_abf2 |
rights_invalid_str_mv |
https://creativecommons.org/licenses/by-nc-nd/4.0/ Atribución-NoComercial-SinDerivadas 4.0 Internacional (CC BY-NC-ND 4.0) http://purl.org/coar/access_right/c_abf2 |
eu_rights_str_mv |
openAccess |
dc.format.extent.spa.fl_str_mv |
10 páginas |
dc.format.mimetype.spa.fl_str_mv |
application/pdf |
dc.coverage.region.none.fl_str_mv |
Antioquia, Colombia |
dc.publisher.spa.fl_str_mv |
Elsevier Science |
dc.publisher.place.spa.fl_str_mv |
Netherlands |
dc.source.spa.fl_str_mv |
https://www.sciencedirect.com/science/article/pii/S1567134821004032 |
institution |
Tecnológico de Antioquia |
bitstream.url.fl_str_mv |
https://dspace.tdea.edu.co/bitstream/tdea/2771/1/Virome%20analysis%20of%20three%20Ixodidae%20ticks%20species%20from%20Colombia_%20A%20potential%20strategy%20for%20discovering%20and%20surveying%20tick-borne%20viruses.pdf https://dspace.tdea.edu.co/bitstream/tdea/2771/2/license.txt https://dspace.tdea.edu.co/bitstream/tdea/2771/3/Virome%20analysis%20of%20three%20Ixodidae%20ticks%20species%20from%20Colombia_%20A%20potential%20strategy%20for%20discovering%20and%20surveying%20tick-borne%20viruses.pdf.txt https://dspace.tdea.edu.co/bitstream/tdea/2771/4/Virome%20analysis%20of%20three%20Ixodidae%20ticks%20species%20from%20Colombia_%20A%20potential%20strategy%20for%20discovering%20and%20surveying%20tick-borne%20viruses.pdf.jpg |
bitstream.checksum.fl_str_mv |
e4777c8f2b53edad1a26eb0cde7202a9 2f9959eaf5b71fae44bbf9ec84150c7a 2c6eb67c8897d916ae47524b1a844d3f 966677552b3546d17280f725b63e693e |
bitstream.checksumAlgorithm.fl_str_mv |
MD5 MD5 MD5 MD5 |
repository.name.fl_str_mv |
Repositorio Institucional Tecnologico de Antioquia |
repository.mail.fl_str_mv |
bdigital@metabiblioteca.com |
_version_ |
1812189204468203520 |
spelling |
Orozco Orozco, Mateoe9774295-d2de-4e2f-8309-75b10e6c1466Gómez García, Giovan Fernandoba1c43f4-0c6e-4605-a4cf-720f02896802Alzate Restrepo, Juan Fernando73945d80-5b97-409d-85c5-59b34c14ac1bIsaza Agudelo, Juan Pablo0b0155c7-8c55-442b-b121-999bed267d78Gutiérrez Builes, Lina Andreae82f4e2a-426b-4f99-b1ac-847f768d54e22023-04-11T19:16:12Z2023-04-11T19:16:12Z20211567-1348https://dspace.tdea.edu.co/handle/tdea/27711567-7257Ticks are a group of obligate blood-sucking ectoparasites that play a critical role in transmitting several important zoonotic pathogens that can infect animals and humans. Viruses are part of the tick microbiome and are involved in the transmission of important diseases. Furthermore, the little information on these as etiological agents of zoonoses suggests the need to study these microorganisms. For this reason, in this study, we sought to characterize the virome in Rhipicephalus microplus, Dermacentor nitens, and Rhipicephalus sanguineus s.l., which were collected from different domestic animals in Antioquia, Colombia. RNA sequencing was used for virome characterization in these three tick species, using RNA-dependent polymerase as a marker gene. Forty-eight sequences corresponding to 14 different viruses were identified, some of which were previously identified in the tick’s virome. Overall, these data indicate that ticks from domestic animals in cattle farms harbor a wide viral diversity at the local scale. Thus, the metatranscriptomic approach provides important baseline information for monitoring the tick virome and to develop future studies on their biology, host-virus interactions, host range, worldwide distribution, and finally, their potential role as emerging vector-borne agents10 páginasapplication/pdfengElsevier ScienceNetherlandshttps://creativecommons.org/licenses/by-nc-nd/4.0/Atribución-NoComercial-SinDerivadas 4.0 Internacional (CC BY-NC-ND 4.0)info:eu-repo/semantics/openAccesshttp://purl.org/coar/access_right/c_abf2https://www.sciencedirect.com/science/article/pii/S1567134821004032Virome analysis of three Ixodidae ticks species from Colombia: A potential strategy for discovering and surveying tick-borne virusesArtículo de revistahttp://purl.org/coar/resource_type/c_2df8fbb1Textinfo:eu-repo/semantics/articlehttp://purl.org/redcol/resource_type/ARTinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/version/c_970fb48d4fbd8a85Antioquia, Colombia10196Infection, genetics and evolutionAdams, M., Kreuze, J., Martelli, G., 2011. Tymovirales [WWW Document]. Int. Comm. Taxon. Viruses ICTVAltschul, S.F., Gish, W., Miller, W., Myers, E.W., Lipman, D.J., 1990. Basic local alignment search tool. J. Mol. Biol. 215, 403–410. https://doi.org/10.1016/S0022- 2836(05)80360-2.Alvarez, V., Bonilla, R., Chac ́on, I., 2003. Frecuencia relativa de Boophilus microplus (Acari: Ixodidae) en bovinos (Bos taurus y B. indicus) en ocho zonas ecol ́ogicas de Costa Rica. Rev. Biol. Trop. 51, 427–434.Andrews, S., 2010. FastQC: A Quality Control Tool for High Throughput Sequence Data [WWW Document]. Online. URL. http://www.bioinformatics.babraham.ac.uk/pro jects/fastqc/.Betancur, O.H., Betancourt, A.E., Giraldo, C.R., 2015. Importance of ticks in the transmission of zoonotic agents Importancia de las garrapatas en la transmisi ́on de agentes causantes de zoonosis. Rev. MVZ C ́ordoba 20, 5053–5067.Bolger, A.M., Lohse, M., Usadel, B., 2014. Trimmomatic: a flexible trimmer for Illumina sequence data. Bioinformatics 30, 2114–2120. https://doi.org/10.1093/ bioinformatics/btu170.Bonnet, S.I., Pollet, T., 2021. Update on the intricate tango between tick microbiomes and tick-borne pathogens. Parasite Immunol. 43, e12813 https://doi.org/10.1111/ pim.12813.Brinkmann, A., Nitsche, A., Kohl, C., 2016. Viral metagenomics on blood-feeding arthropods as a tool for human disease surveillance. Int. J. Mol. Sci. 17, E1743 https://doi.org/10.3390/ijms17101743.Buchfink, B., Xie, C., Huson, D.H., 2015. Fast and sensitive protein alignment using DIAMOND. Nat. Methods 12, 59–60. https://doi.org/10.1038/nmeth.3176.C ́amara de Comercio de Medellín para Antioquia, 2017. Perfil Socioecon ́omico de la Subregi ́on Magdalena Medio [WWW Document]. Inf. Estud. Econ ́omicos. URL https ://www.camaramedellin.com.co/Portals/0/Biblioteca/Estudios-economicos/cade nas-productivas-regionales/12-3Perfil Magdalena_Oct14.pdf?ver=2019-03-01- 095037-233 (accessed 3.3.21).Colombo, V.C., Fasano, A.A., Beldomenico, P.M., Nava, S., 2018. Tick host specificity: an analysis based on host phylogeny and tick ecological features using Amblyomma triste and Amblyomma tigrinum immature stages. Ticks Tick. Borne. Dis. 9, 781–787. https://doi.org/10.1016/j.ttbdis.2018.03.001.Cross, S.T., Kapuscinski, M.L., Perino, J., Maertens, B.L., Weger-Lucarelli, J., Ebel, G.D., Stenglein, M.D., 2018. Co-infection patterns in individual Ixodes scapularis ticks reveal associations between viral, eukaryotic and bacterial microorganisms. Viruses 10, 388. https://doi.org/10.3390/v10070388.de Souza, W.M., Fumagalli, M.J., Carrasco, Torres, Romeiro, M.F., Modha, S., Seki, M.C., Gheller, J.M., Daffre, S., Nunes, M.R.T., Murcia, P.R., Acrani, G.O., Figueiredo, L.T. M., 2018. Viral diversity of Rhipicephalus microplus parasitizing cattle in southern Brazil. Sci. Rep. 8, 16315. https://doi.org/10.1038/s41598-018-34630-1.Easton, A.J., Pringle, C.R., 2011. Mononegavirales [WWW Document]. Noveno Inf. ICTV. Eddy, S.R., 2011. Accelerated profile HMM searches. PLoS Comput. Biol. 7. https://doi. org/10.1371/journal.pcbi.1002195.Edgar, R.C., 2004. MUSCLE: multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Res. 32, 1792–1797. https://doi.org/10.1093/nar/ gkh340.Esser, H.J., Herre, E.A., Kays, R., Liefting, Y., Jansen, P.A., 2019. Local host-tick coextinction in neotropical forest fragments. Int. J. Parasitol. 49, 225–233. https:// doi.org/10.1016/j.ijpara.2018.08.008.Estrada-Pe ̃na, A., 2008. Climate, niche, ticks, and models: what they are and how we should interpret them. Parasitol. Res. 103, S87–S95. https://doi.org/10.1007/ s00436-008-1056-7Estrada-Pe ̃na, A., Nava, S., Tarragona, E., de la Fuente, J., Guglielmone, A.A., 2020. A community approach to the Neotropical ticks-hosts interactions. Sci. Rep. 10, 9269. https://doi.org/10.1038/s41598-020-66400-3.Faizah, A.N., Kobayashi, D., Isawa, H., Amoa-Bosompem, M., Murota, K., Higa, Y., Futami, K., Shimada, S., Kim, K.S., Itokawa, K., Watanabe, M., Tsuda, Y., Minakawa, N., Miura, K., Hirayama, K., Sawabe, K., 2020. Deciphering the virome of Culex vishnui subgroup mosquitoes, the major vectors of japanese encephalitis, in Japan. Viruses 12, 264. https://doi.org/10.3390/v12030264.G ́omez, G.F., Isaza, J.P., Segura, J.A., Alzate, J.F., Guti ́errez, L.A., 2020. Metatranscriptomic virome assessment of Rhipicephalus microplus from Colombia. Ticks Tick. Borne. Dis. 11, 101426. https://doi.org/10.1016/j.ttbdis.2020.101426.Gondard, M., Temmam, S., Devillers, E., Bigot, T., Chr, D., Aprelon, R., Vayssier- taussat, M., Albina, E., Eloit, M., Moutailler, S., 2020. RNA Viruses of Amblyomma Variegatum and Rhipicephalus Microplus and Cattle Susceptibility in the French Antilles, pp. 1–23Grabherr, M.G., Haas, B.J., Yassour, M., Levin, J.Z., Thompson, D.A., Amit, I., Adiconis, X., Fan, L., Raychowdhury, R., Zeng, Q., Chen, Z., Mauceli, E., Hacohen, N., Gnirke, A., Rhind, N., di Palma, F., Birren, B.W., Nusbaum, C., Lindblad Toh, K., Friedman, N., Regev, A., 2013. Trinity: reconstructing a full-length transcriptome without a genome from RNA-Seq data. Nat. Biotechnol. 29, 644–652. https://doi.org/10.1038/nbt.1883.TrinityGrard, G., Moureau, G., Charrel, R.N., Lemasson, J.J., Gonzalez, J.P., Gallian, P., Gritsun, T.S., Holmes, E.C., Gould, E.A., de Lamballerie, X., 2007. Genetic characterization of tick-borne flaviviruses: new insights into evolution, pathogenetic determinants and taxonomy. Virology 361, 80–92. https://doi.org/10.1016/j. virol.2006.09.015.Guglielmone, A.A., Apanaskevich, D.A., Estrada-Pe ̃na, A., Robbins, R.G., Petney, T.N., Horak, I.G., 2014. The hard ticks of the world: (Acari: Ixodida: Ixodidae). In: The Hard Ticks of the World: (Acari: Ixodida: Ixodidae). Springer Netherlands, Dordrecht, The Netherlands. https://doi.org/10.1007/978-94-007-7497-1.Harris, H.M.B., Hill, C., 2021. A place for viruses on the tree of life. Front. Microbiol. 11, 604048. https://doi.org/10.3389/fmicb.2020.604048.Huson, D.H., Auch, A.F., Qi, J., Schuster, S.C., 2007. MEGAN analysis of metagenomic data. Genome Res. 17, 377–386. https://doi.org/10.1101/gr.5969107.Hyatt, D., Locascio, P.F., Hauser, L.J., Uberbacher, E.C., 2012. Gene and translation initiation site prediction in metagenomic sequences. Bioinformatics 28, 2223–2230. https://doi.org/10.1093/bioinformatics/bts429.International Committee on Taxonomy of Viruses, 2012. Tymoviridae. In: King, A.M.Q., Adams, M.J., Carstens, E.B., Lefkowitz, E.J. (Eds.), Virus Taxonomy. Elsevier, pp. 944–952. https://doi.org/10.1016/b978-0-12-384684-6.00080-x.Kazimírov ́a, M., Thangamani, S., Bartíkov ́a, P., Hermance, M., Holíkov ́a, V., ˇStibr ́aniov ́a, I., Nuttall, P.A., 2017. Tick-borne viruses and biological processes at the tick-host-virus interface. Front. Cell. Infect. Microbiol. 7, 1–21. https://doi.org/ 10.3389/fcimb.2017.00339.Kazimírov ́a, M., Bartíkov ́a, P., ˇStibr ́aniov ́a, I., 2018. Tick-borne viruses and host skin interface. In: Skin and Arthropod Vectors. Elsevier Inc, pp. 325–383. https://doi.org/ 10.1016/B978-0-12-811436-0.00010-1.Kumar, A., Murthy, S., Kapoor, A., 2017. Evolution of selective-sequencing approaches for virus discovery and virome analysis. Virus Res. https://doi.org/10.1016/j. virusres.2017.06.005.Letunic, I., Bork, P., 2021. Interactive tree of life (iTOL) v5: an online tool for phylogenetic tree display and annotation. Nucleic Acids Res. https://doi.org/ 10.1093/nar/gkab301 gkab301Li, H., Durbin, R., 2009. Fast and accurate short read alignment with burrows-wheeler transform. Bioinformatics 25, 1754–1760. https://doi.org/10.1093/bioinformatics/ btp324.Li, C.X., Shi, M., Tian, J.H., Lin, X.D., Kang, Y.J., Chen, L.J., Qin, X.C., Xu, J., Holmes, E. C., Zhang, Y.Z., 2015. Unprecedented genomic diversity of RNA viruses in arthropods reveals the ancestry of negative-sense RNA viruses. Elife 4, e05378. https://doi.org/10.7554/eLife.05378.Liu, Q., He, B., Huang, S.Y., Wei, F., Zhu, X.Q., 2014. Severe fever with thrombocytopenia syndrome, an emerging tick-borne zoonosis. Lancet Infect. Dis. 14, 763–772. https://doi.org/10.1016/S1473-3099(14)70718-2.L ́opez, Y., Miranda, J., Mattar, S., Gonzalez, M., Rovnak, J., 2020. First report of Lihan Tick virus (Phlebovirus, Phenuiviridae) in ticks, Colombia. Virol. J. 17, 1–6. https:// doi.org/10.1186/s12985-020-01327-9.Madison-Antenucci, S., Kramer, L.D., Gebhardt, L.L., Kauffman, E., 2020. Emerging tick- borne diseases. Clin. Microbiol. Rev. https://doi.org/10.1128/CMR e00083–18. Martelli, G.P., Adams, M.J., Kreuze, J.F., Dolja, V.V., 2008. Family Flexiviridae: a case study in virion and genome plasticity. Annu. Rev. Phytopathol. 45, 73–100. https:// doi.org/10.1146/annurev.phyto.45.062806.094401.Meng, F., Ding, M., Tan, Z., Zhao, Z., Xu, L., Wu, J., He, B., Tu, C., 2018. Virome analysis of tick-borne viruses in Heilongjiang Province, China. Ticks Tick. Borne. Dis. 10, 412–420. https://doi.org/10.1016/j.ttbdis.2018.12.002.Monsalve-Arteagaid, L., Alonso-Sard ́on, M., Bellido, J.L.M., Santiago, M.B.V., Lista, M.C. V., Ab ́an, J.L., Muro, A., Belhassen-García, M., 2020. Seroprevalence of crimean- congo hemorrhagic fever in humans in the world health organization european region: a systematic review. PLoS Negl. Trop. Dis. 14, e0008094 https://doi.org/ 10.1371/journal.pntd.0008094.Morrone, J.J., 2014. Biogeographical regionalisation of the Neotropical region. Zootaxa 3782, 1–110. https://doi.org/10.11646/zootaxa.3782.1.1.Narasimhan, S., Fikrig, E., 2015. Tick microbiome: the force within. Trends Parasitol. 31, 315–323. https://doi.org/10.1016/j.pt.2015.03.010.Nava, S., Guglielmone, A.A., 2013. A meta-analysis of host specificity in Neotropical hard ticks (Acari: Ixodidae). Bull. Entomol. Res. 103, 216–224. https://doi.org/10.1017/ S0007485312000557Ortíz-Giraldo, M., Tob ́on-Escobar, W.D., Vel ́asquez-Guarín, D., Usma-Marín, M.F., Ossa- L ́opez, P.A., Ramírez-Chaves, H.E., Carvajal-Agudelo, J.D., Rivera-P ́aez, F.A., 2021. Ticks (Acari: Ixodoidea) associated with mammals in Colombia: a historical review, molecular species confirmation, and establishment of new relationships. Parasitol. Res. 120, 383–394. https://doi.org/10.1007/s00436-020-06989-6.Paules, C.I., Marston, H.D., Bloom, M.E., Fauci, A.S., 2018. Tickborne diseases — confronting a growing threat. N. Engl. J. Med. 379, 701–703. https://doi.org/ 10.1056/nejmp1807870.Prasad, N., Murdoch, D.R., Reyburn, H., Crump, J.A., 2015. Etiology of severe febrile illness in low- and middle-income countries: a systematic review. PLoS One 10, e0127962. https://doi.org/10.1371/journal.pone.0127962Regier, Y., Komma, K., Weigel, M., Kraiczy, P., Laisi, A., Pulliainen, A.T., Hain, T., Kempf, V.A.J., 2019. Combination of microbiome analysis and serodiagnostics to assess the risk of pathogen transmission by ticks to humans and animals in Central Germany 11 medical and health sciences 1108 medical microbiology. Parasit. Vectors 12. https://doi.org/10.1186/s13071-018-3240-7Rosenberg, R., Lindsey, N.P., Fischer, M., Gregory, C.J., Hinckley, A.F., Mead, P.S., Paz- Bailey, G., Waterman, S.H., Drexler, N.A., Kersh, G.J., Hooks, H., Partridge, S.K., Visser, S.N., Beard, C.B., Petersen, L.R., 2018. Vital signs: trends in reported vectorborne disease cases — United States and territories, 2004–2016. MMWR Morb. Mortal. Wkly Rep. 67, 496–501. https://doi.org/10.15585/mmwr. mm6717e1.Sameroff, S., Tokarz, R., Charles, R.A., Jain, K., Oleynik, A., Che, X., Georges, K., Carrington, C.V., Lipkin, W.I., Oura, C., 2019. Viral diversity of tick species parasitizing cattle and dogs in Trinidad and Tobago. Sci. Rep. 9, 10421. https://doi. org/10.1038/s41598-019-46914-1.Savage, H.M., Godsey, M.S., Amy, L., Panella, N.A., Burkhalter, K.L., Harmon, J.R., Lash, R.R., Ashley, D.C., Nicholson, W.L., 2013. First detection of Heartland virus (bunyaviridae: Phlebovirus) from field collected arthropods. Am. J. Trop. Med. Hyg. 89, 445–452. https://doi.org/10.4269/ajtmh.13-0209.. Segura, J.A., Isaza, J.P., Botero, L.E., Alzate, J.F., Guti ́errez, L.A., 2020. Assessment of bacterial diversity of Rhipicephalus microplus ticks from two livestock agroecosystems in Antioquia, Colombia. PLoS One 15. https://doi.org/10.1371/ journal.pone.0234005.Shi, M., Lin, X.D., Tian, J.H., Chen, L.J., Chen, X., Li, C.X., Qin, X.C., Li, J., Cao, J.P., Eden, J.S., Buchmann, J., Wang, W., Xu, J., Holmes, E.C., Zhang, Y.Z., 2016. Redefining the invertebrate RNA virosphere. Nature 540, 539–543. https://doi.org/ 10.1038/nature20167Shi, J., Hu, Z., Deng, F., Shen, S., 2018. Tick-borne viruses. Virol. Sin. 33, 21–43. https:// doi.org/10.1007/s12250-018-0019-0.Shi, J., Shen, S., Wu, H., Zhang, Y., Deng, F., 2021. Metagenomic profiling of viruses associated with Rhipicephalus microplus ticks in Yunnan Province, China. Virol. Sin. https://doi.org/10.1007/s12250-020-00319-x.Stamatakis, A., 2014. RAxML version 8: a tool for phylogenetic analysis and post-analysis of large phylogenies. Bioinformatics 30, 1312–1313. https://doi.org/10.1093/ bioinformatics/btu033.Segura, J.A., Isaza, J.P., Botero, L.E., Alzate, J.F., Guti ́errez, L.A., 2020. Assessment of bacterial diversity of Rhipicephalus microplus ticks from two livestock agroecosystems in Antioquia, Colombia. PLoS One 15. https://doi.org/10.1371/ journal.pone.0234005Starr, E.P., Nuccio, E.E., Pett-Ridge, J., Banfield, J.F., Firestone, M.K., 2019. Metatranscriptomic reconstruction reveals RNA viruses with the potential to shape carbon cycling in soil. bioRxiv 116, 25900–25908. https://doi.org/10.1101/ 597468.Temmam, S., Chr ́etien, D., Bigot, T., Dufour, E., Petres, S., Desquesnes, M., Devillers, E., Dumarest, M., Yousfi, L., Jittapalapong, S., Karnchanabanthoeng, A., Chaisiri, K., Gagnieur, L., Cosson, J.F., Vayssier-Taussat, M., Morand, S., Moutailler, S., Eloit, M., 2019. Monitoring silent spillovers before emergence: a pilot study at the tick/human interface in Thailand. Front. Microbiol. 10, 2315. https://doi.org/10.3389/ fmicb.2019.02315.Tokarz, R., Williams, S.H., Sameroff, S., Sanchez Leon, M., Jain, K., Lipkin, W.I., 2014. Virome analysis of Amblyomma americanum, Dermacentor variabilis, and Ixodes scapularis ticks reveals novel highly divergent vertebrate and invertebrate viruses. J. Virol. 88, 11480–11492. https://doi.org/10.1128/JVI.01858-14.Vandegrift, K.J., Kapoor, A., 2019. The ecology of new constituents of the tick virome and their relevance to public health. Viruses 11, 529. https://doi.org/10.3390/ v11060529.Walker, A.R., Bouattour, A., Camicas, J., Estrada-Pe ̃na, A., Horak, I.G., Latif, A.A., Pegram, R.G., Preston, P.M., 2003. Ticks of Domestic Animals in Africa: A Guide to Identification of Species. Bioscience Reports, Edinburgh Scotland,U.K.Wang, L., Lv, X., Zhai, Y., Fu, S., Wang, D., Rayner, S., Tang, Q., Liang, G., 2012. Genomic characterization of a novel virus of the family Tymoviridae isolated from mosquitoes. PLoS One 7. https://doi.org/10.1371/journal.pone.0039845.Waterhouse, R.M., Seppey, M., Simao, F.A., Manni, M., Ioannidis, P., Klioutchnikov, G., Kriventseva, E.V., Zdobnov, E., 2018. BUSCO applications from quality assessments to gene prediction and phylogenomics. Mol. Biol. Evol. 35, 543–548. https://doi. org/10.1093/molbev/msx319Wolf, Y.I., Kazlauskas, D., Iranzo, J., Luc ́Ia-Sanz, A., Kuhn, J.H., Krupovic, M., Dolja, V. V., Koonin, E.V., 2018. Origins and evolution of the global RNA virome. MBio 9, 1–31. https://doi.org/10.1101/451740. Wu, H., Pang, R., Cheng, T., Xue, L., Zeng, H., Lei, T., Chen, M., Wu, S., Ding, Y., Zhang, J., Shi, M., Wu, Q., 2020. Abundant and diverse RNA viruses in insects revealed by RNA-Seq analysis: ecological and evolutionary implications. mSystems 5. https://doi.org/10.1128/msystems.00039-20.Zhao, T., Gong, H., Shen, X., Zhang, W., Shan, T., Yu, X., Wang, S.J., Cui, L., 2020. Comparison of viromes in ticks from fifferent domestic animals in China. Virol. Sin. 35, 398–406. https://doi.org/10.1007/s12250-020-00197-3.RhipicephalusRhipicephalus sanguineusGarrapata Marrón del PerroZoonosisTick-borne diseasesEnfermedades por Picaduras de GarrapatasDoenças Transmitidas por CarrapatosRhipicephalus microplusDermacentor nitensORIGINALVirome analysis of three Ixodidae ticks species from Colombia_ A potential strategy for discovering and surveying tick-borne viruses.pdfVirome analysis of three Ixodidae ticks species from Colombia_ A potential strategy for discovering and surveying tick-borne viruses.pdfapplication/pdf6590633https://dspace.tdea.edu.co/bitstream/tdea/2771/1/Virome%20analysis%20of%20three%20Ixodidae%20ticks%20species%20from%20Colombia_%20A%20potential%20strategy%20for%20discovering%20and%20surveying%20tick-borne%20viruses.pdfe4777c8f2b53edad1a26eb0cde7202a9MD51open accessLICENSElicense.txtlicense.txttext/plain; charset=utf-814828https://dspace.tdea.edu.co/bitstream/tdea/2771/2/license.txt2f9959eaf5b71fae44bbf9ec84150c7aMD52open accessTEXTVirome analysis of three Ixodidae ticks species from Colombia_ A potential strategy for discovering and surveying tick-borne viruses.pdf.txtVirome analysis of three Ixodidae ticks species from Colombia_ A potential strategy for discovering and surveying tick-borne viruses.pdf.txtExtracted texttext/plain10https://dspace.tdea.edu.co/bitstream/tdea/2771/3/Virome%20analysis%20of%20three%20Ixodidae%20ticks%20species%20from%20Colombia_%20A%20potential%20strategy%20for%20discovering%20and%20surveying%20tick-borne%20viruses.pdf.txt2c6eb67c8897d916ae47524b1a844d3fMD53open accessTHUMBNAILVirome analysis of three Ixodidae ticks species from Colombia_ A potential strategy for discovering and surveying tick-borne viruses.pdf.jpgVirome analysis of three Ixodidae ticks species from Colombia_ A potential strategy for discovering and surveying tick-borne viruses.pdf.jpgGenerated Thumbnailimage/jpeg15160https://dspace.tdea.edu.co/bitstream/tdea/2771/4/Virome%20analysis%20of%20three%20Ixodidae%20ticks%20species%20from%20Colombia_%20A%20potential%20strategy%20for%20discovering%20and%20surveying%20tick-borne%20viruses.pdf.jpg966677552b3546d17280f725b63e693eMD54open accesstdea/2771oai:dspace.tdea.edu.co:tdea/27712023-04-12 03:01:16.635An error occurred on the license name.|||https://creativecommons.org/licenses/by-nc-nd/4.0/open accessRepositorio Institucional Tecnologico de Antioquiabdigital@metabiblioteca.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 |