{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,17]],"date-time":"2026-07-17T15:30:40Z","timestamp":1784302240281,"version":"3.55.0"},"reference-count":86,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2024,12,8]],"date-time":"2024-12-08T00:00:00Z","timestamp":1733616000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100004587","name":"Instituto de Salud Carlos III","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100004587","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100013239","name":"Centro de Estudos Ambientais e Marinhos, Universidade de Aveiro","doi-asserted-by":"publisher","award":["UIDP\/50017\/2020\u2009+\u2009UIDB\/50017\/2020\u2009+\u2009LA\/P\/0094\/2020"],"award-info":[{"award-number":["UIDP\/50017\/2020\u2009+\u2009UIDB\/50017\/2020\u2009+\u2009LA\/P\/0094\/2020"]}],"id":[{"id":"10.13039\/100013239","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004587","name":"Instituto de Salud Carlos III","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100004587","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Zoonoses and Public Health"],"published-print":{"date-parts":[[2025,5]]},"abstract":"<jats:title>ABSTRACT<\/jats:title><jats:sec><jats:title>Introduction<\/jats:title><jats:p>Intestinal microeukaryote parasites are major contributors to the burden of diarrhoea in humans and domestic animals, but their epidemiology in wildlife is not fully understood. We investigated the frequency, genetic diversity and zoonotic potential of protists of animal and public health significance in free\u2010ranging grey wolf (<jats:italic>Canis lupus<\/jats:italic>) populations in south\u2010western Europe.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Individually formed faecal samples collected from necropsied wolves or scat trails in Italy (<jats:italic>n<\/jats:italic>\u2009=\u200947), Portugal (<jats:italic>n<\/jats:italic>\u2009=\u200943) and Spain (<jats:italic>n<\/jats:italic>\u2009=\u2009225) during the period 2011\u20132023 were retrospectively analysed using molecular (PCR and Sanger sequencing) methods. Complementary epidemiological data were gathered when available.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>\n<jats:italic>Giardia duodenalis<\/jats:italic> was the most frequent microeukaryote found (40.3%, 127\/315; 95% CI: 34.9\u201346.0), followed by <jats:italic>Cryptosporidium<\/jats:italic> spp. (3.5%, 11\/315; 95% CI: 1.8\u20136.2), <jats:italic>Enterocytozoon bieneusi<\/jats:italic> and <jats:italic>Encephalitozoon<\/jats:italic> spp. (1.6%, 5\/315; 95% CI: 0.5\u20133.7 each). <jats:italic>Blastocystis<\/jats:italic> was not identified in any of the faecal samples analysed. Sequence analyses confirmed the presence of canine\u2010adapted assemblage D within <jats:italic>G. duodenalis<\/jats:italic> (<jats:italic>n<\/jats:italic>\u2009=\u20097). Three <jats:italic>Cryptosporidium<\/jats:italic> species were identified, namely canine\u2010adapted <jats:italic>C. canis<\/jats:italic> (<jats:italic>n<\/jats:italic>\u2009=\u20099), zoonotic <jats:italic>C.\u2009parvum<\/jats:italic> (<jats:italic>n<\/jats:italic>\u2009=\u20091) and primarily anthroponotic <jats:italic>C.hominis<\/jats:italic> (<jats:italic>n<\/jats:italic>\u2009=\u20091). Genotyping tools enabled the identification of subtype family XXe2 within <jats:italic>C. canis<\/jats:italic>. Among microsporidia, the canine\u2010adapted genotype PtEb IX was identified within <jats:italic>E. bieneusi<\/jats:italic>. Two samples were confirmed as <jats:italic>Enc. intestinalis<\/jats:italic> and three more as <jats:italic>Enc. cuniculi<\/jats:italic> genotype IV. This is the first record of <jats:italic>Enc. intestinalis<\/jats:italic> and <jats:italic>Enc. cuniculi<\/jats:italic> in the grey wolf globally.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>Silent carriage of intestinal microeukaryotes seems common in free\u2010ranging grey wolves in southwestern Europe. Wolves can contribute to environmental contamination through the transmission stages (cysts, oocysts, spores) of species\/genotypes potentially infective to humans. Individuals in close contact with wolf carcasses or their faecal material may be at potential risk of infection by microeukaryotic pathogens.<\/jats:p><\/jats:sec>","DOI":"10.1111\/zph.13202","type":"journal-article","created":{"date-parts":[[2024,12,9]],"date-time":"2024-12-09T07:29:02Z","timestamp":1733729342000},"page":"269-283","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Free\u2010Ranging Wolves (<i>Canis lupus<\/i>) are Natural Reservoirs of Intestinal Microeukaryotes of Public Health Significance in Southwestern Europe"],"prefix":"10.1111","volume":"72","author":[{"given":"Sheila","family":"Ortega","sequence":"first","affiliation":[{"name":"Parasitology Reference and Research Laboratory National Centre for Microbiology  Majadahonda Madrid Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2623-6340","authenticated-orcid":false,"given":"Ana M.","family":"Figueiredo","sequence":"additional","affiliation":[{"name":"CESAM and Department of Biology University of Aveiro  Aveiro Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Barbara","family":"Moroni","sequence":"additional","affiliation":[{"name":"Istituto Zooprofilattico di Piemonte, Liguria e Valle d'Aosta (IZSPLV)  Torino Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nadia","family":"Abarca","sequence":"additional","affiliation":[{"name":"Parasitology Reference and Research Laboratory National Centre for Microbiology  Majadahonda Madrid Spain"},{"name":"Department of Veterinary Sciences, Biomedical Sciences Institute Autonomous University of Ciudad Ju\u00e1rez  Chihuahua Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alejandro","family":"Dashti","sequence":"additional","affiliation":[{"name":"Parasitology Reference and Research Laboratory National Centre for Microbiology  Majadahonda Madrid Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pamela C.","family":"K\u00f6ster","sequence":"additional","affiliation":[{"name":"Parasitology Reference and Research Laboratory National Centre for Microbiology  Majadahonda Madrid Spain"},{"name":"Faculty of Health Sciences Alfonso X El Sabio University (UAX)  Villanueva de la Ca\u00f1ada, Madrid Spain"},{"name":"Women for Africa Foundation  Madrid Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bego\u00f1a","family":"Bailo","sequence":"additional","affiliation":[{"name":"Parasitology Reference and Research Laboratory National Centre for Microbiology  Majadahonda Madrid Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David","family":"Cano\u2010Terriza","sequence":"additional","affiliation":[{"name":"Department of Animal Health, Animal Health and Zoonoses Research Group (GISAZ), UIC Zoonoses and Emerging Diseases (ENZOEM) University of C\u00f3rdoba  C\u00f3rdoba Spain"},{"name":"CIBER Infectious Diseases (CIBERINFEC), Health Institute Carlos III  Madrid Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0423-3189","authenticated-orcid":false,"given":"Mois\u00e9s","family":"Gonz\u00e1lvez","sequence":"additional","affiliation":[{"name":"Department of Animal Health, Animal Health and Zoonoses Research Group (GISAZ), UIC Zoonoses and Emerging Diseases (ENZOEM) University of C\u00f3rdoba  C\u00f3rdoba Spain"},{"name":"Department of Animal Health, Faculty of Veterinary Sciences, Regional Campus of International Excellence \u201cCampus Mare Nostrum\u201d University of Murcia  Murcia Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Manena","family":"Fayos","sequence":"additional","affiliation":[{"name":"Centro de Recuperaci\u00f3n de Fauna Silvestre de Cantabria, Tragsatec, Direcci\u00f3n General de Montes y Biodiversidad Cantabria Gobierno de Cantabria  Santander Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"\u00c1lvaro","family":"Oleaga","sequence":"additional","affiliation":[{"name":"Sociedad de Servicios del Principado de Asturias S.A. 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