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The emergence of highly pathogenic PRRS virus 1 (PRRSV\u20101) strains, first detected in northeastern Spain in 2020, poses new challenges for effective disease control. Despite their severe impact under field conditions, a comprehensive experimental characterization of these virulent strains has been lacking. In this study, we analyzed the genetic characteristics of the Lleida 029_22 strain, collected during a severe PRRSV\u20101 outbreak, and evaluated its pathogenicity in pigs. Phylogenetic analysis revealed that the Lleida 029_22 strain belongs to a new clade describing multiple strains isolated during a PRRSV\u20101 Rosalia\u2010related outbreak, with the highly pathogenic Italian strain PR40 identified as a common ancestor.\n                    <jats:italic>In vitro<\/jats:italic>\n                    , this strain replicated efficiently in porcine alveolar macrophages and PAM\u2010KNU cells, but not in MARC\u2010145 cells. To characterize the pathogenicity of this strain, 8\u2010week\u2010old piglets were inoculated with a 2\u2009\u00d7\u200910\n                    <jats:sup>5<\/jats:sup>\n                    TCID\n                    <jats:sub>50<\/jats:sub>\n                    dose of the Lleida 029_22 strain via two inoculation routes\u2014intramuscular (IM) and intranasal (IN). The IM\u2010infected pigs exhibited high levels of viremia and virus shedding, with 100% mortality recorded by 14 days post\u2010infection (dpi). These animals also showed significantly elevated levels of pro\u2010inflammatory cytokines, particularly IL\u20106, and severe histopathologic lung lesions, correlating with the observed severe disease outcome. In contrast, IN\u2010infected pigs exhibited lower mortality (30%) and moderate clinical signs, with the surviving pigs fully recovering by the end of the 63\u2010day observation period. These pigs also exhibited prolonged viremia and viral shedding, with lower levels of pro\u2010inflammatory cytokines and neutralizing antibodies from 28\u2009dpi. The IN infection closely mimicked the disease outcomes observed in Spanish farms experiencing outbreaks due to the high\u2010virulence Rosalia strain, while the IM infection highlighted the risks of iatrogenic transmission.\n                  <\/jats:p>","DOI":"10.1155\/tbed\/4847981","type":"journal-article","created":{"date-parts":[[2025,5,23]],"date-time":"2025-05-23T07:19:22Z","timestamp":1747984762000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["A Spanish Porcine Reproductive and Respiratory Syndrome Virus 1 Strain Is Highly Virulent in Pigs Under Experimental Conditions"],"prefix":"10.1155","volume":"2025","author":[{"given":"Paula","family":"Curto","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ariadna","family":"Pey","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alba","family":"Trabal","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elisabet","family":"Aulinas","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jordi","family":"Baliellas","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gustavo","family":"Ram\u00edrez","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"J\u00e9ssica","family":"Mol\u00edn","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Larysa","family":"Muzykina","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria","family":"Montoya","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ramona N.","family":"Pena","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lorenzo","family":"Fraile","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6687-7134","authenticated-orcid":false,"given":"Ana M. 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