{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,31]],"date-time":"2026-01-31T11:51:53Z","timestamp":1769860313780,"version":"3.49.0"},"reference-count":40,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2025,4,2]],"date-time":"2025-04-02T00:00:00Z","timestamp":1743552000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"RumiRes\u201d, National Funds by FCT\u2014Portuguese Foundation for Science and Technology","award":["PRR-C05-i03-I-000190"],"award-info":[{"award-number":["PRR-C05-i03-I-000190"]}]},{"name":"RumiRes\u201d, National Funds by FCT\u2014Portuguese Foundation for Science and Technology","award":["CERNAS UIDB\/00681\/2020"],"award-info":[{"award-number":["CERNAS UIDB\/00681\/2020"]}]},{"name":"RumiRes\u201d, National Funds by FCT\u2014Portuguese Foundation for Science and Technology","award":["CITAB UIDB\/04033\/2020"],"award-info":[{"award-number":["CITAB UIDB\/04033\/2020"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Veterinary Sciences"],"abstract":"<jats:p>Research on endoparasites in pet dogs has been growing, but shepherd dogs have largely been overlooked. These dogs frequently share close proximity not only with sheep, which are reservoirs of zoonotic subtypes of Blastocystis, but also with their owners. This close contact increases the potential for shepherd dogs to act as intermediates in the transmission of Blastocystis. To clarify the role of these dogs as reservoirs for this parasite, this study investigated the presence of Blastocystis in shepherd dogs. Stool samples from Portuguese shepherd dogs were analyzed using SYBR-Green-based real-time PCR and melting curve analysis followed by targeted-amplicon NGS for mixed infections detection. Our results revealed a 60% occurrence of Blastocystis sp. in shepherd dog stools and frequent identification of zoonotic subtypes ST1\u2013ST4 and ST14. Additionally, we observed mixed infections and subtype diversity within individual dogs, suggesting a potential role in cross-species transmission between livestock and humans.<\/jats:p>","DOI":"10.3390\/vetsci12040325","type":"journal-article","created":{"date-parts":[[2025,4,2]],"date-time":"2025-04-02T04:06:05Z","timestamp":1743566765000},"page":"325","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Targeted-Amplicon NGS for Blastocystis sp. in Shepherd Dogs of Portugal Discriminates Co-Colonization with Multiple Zoonotic Subtypes"],"prefix":"10.3390","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8374-3739","authenticated-orcid":false,"given":"Sara","family":"Gomes-Gon\u00e7alves","sequence":"first","affiliation":[{"name":"School of Medicine and Biomedical Sciences (ICBAS), Universidade do Porto (UP), 4050-313 Porto, Portugal"}]},{"given":"Maria Jo\u00e3o","family":"Feiteiro","sequence":"additional","affiliation":[{"name":"Instituto Polit\u00e9cnico de Viseu, Escola Superior Agr\u00e1ria de Viseu, Campus Polit\u00e9cnico, 3504-510 Viseu, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-2828-7202","authenticated-orcid":false,"given":"Guilherme","family":"Moreira","sequence":"additional","affiliation":[{"name":"School of Medicine and Biomedical Sciences (ICBAS), Universidade do Porto (UP), 4050-313 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5767-7835","authenticated-orcid":false,"given":"Rita","family":"Cruz","sequence":"additional","affiliation":[{"name":"Instituto Polit\u00e9cnico de Viseu, Escola Superior Agr\u00e1ria de Viseu, Campus Polit\u00e9cnico, 3504-510 Viseu, Portugal"},{"name":"Epidemiology Research Unit (EPIUnit), Instituto de Sa\u00fade P\u00fablica, Universidade do Porto (UP), 4050-600 Porto, Portugal"}]},{"given":"Fernando","family":"Esteves","sequence":"additional","affiliation":[{"name":"Instituto Polit\u00e9cnico de Viseu, Escola Superior Agr\u00e1ria de Viseu, Campus Polit\u00e9cnico, 3504-510 Viseu, Portugal"},{"name":"CERNAS-IPV Research Centre, Instituto Polit\u00e9cnico de Viseu, Campus Polit\u00e9cnico, Repeses, 3504-510 Viseu, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6829-4867","authenticated-orcid":false,"given":"Helena","family":"Vala","sequence":"additional","affiliation":[{"name":"Instituto Polit\u00e9cnico de Viseu, Escola Superior Agr\u00e1ria de Viseu, Campus Polit\u00e9cnico, 3504-510 Viseu, Portugal"},{"name":"CERNAS-IPV Research Centre, Instituto Polit\u00e9cnico de Viseu, Campus Polit\u00e9cnico, Repeses, 3504-510 Viseu, Portugal"},{"name":"Centre for the Research and Technology of Agro-Environmental and Biological Sciences (CITAB), University of Tr\u00e1s-os-Montes e Alto Douro, 5001-801 Vila Real, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8769-8103","authenticated-orcid":false,"given":"Jo\u00e3o R.","family":"Mesquita","sequence":"additional","affiliation":[{"name":"School of Medicine and Biomedical Sciences (ICBAS), Universidade do Porto (UP), 4050-313 Porto, Portugal"},{"name":"Associate Laboratory for Animal and Veterinary Science (AL4AnimalS), 1300-477 Lisboa, Portugal"},{"name":"Centro de Estudos de Ci\u00eancia Animal (CECA), Instituto de Ci\u00eancias, Tecnologias e Agroambiente (ICETA), Universidade do Porto (UP), Rua D. Manuel II, Apartado 55142, 4051-401 Porto, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2025,4,2]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"523","DOI":"10.1016\/j.pt.2013.08.006","article-title":"Blastocystis: Getting to grips with our guileful guest","volume":"29","author":"Scanlan","year":"2013","journal-title":"Trends Parasitol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"106377","DOI":"10.1016\/j.micpath.2023.106377","article-title":"Prevalence and genetic diversity of Blastocystis sp. among autochthonous and immigrant patients in Italy","volume":"185","author":"Marangi","year":"2023","journal-title":"Microb. Pathog."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"260","DOI":"10.1016\/j.rvsc.2020.09.031","article-title":"Blastocystis in domesticated and wild mammals and birds","volume":"135","author":"Hublin","year":"2021","journal-title":"Res. Vet. 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