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Italy is one of the few European countries where the circulation of <jats:italic>Besnoitia<\/jats:italic> spp. antibodies was demonstrated. In this study, a case of clinical besnoitiosis in two donkeys in northern Italy is reported. The two animals were clinically examined. Serum and blood samples were analyzed for the detection of <jats:italic>Besnoitia<\/jats:italic> spp. antibodies and for hematology, biochemistry, and enzyme activity, respectively. ITS-1 PCR and sequencing were carried out on DNA extracted from skin biopsies. Clinical examination revealed numerous scleral pearls in eyes of both animals; alopecia and hyperkeratosis with skin nodules in the region of the neck, hind leg, and on the pinnae were detected. No cysts were evidenced by endoscopy in respiratory and genital tracts. Both animals resulted seropositive to <jats:italic>Besnoitia<\/jats:italic> spp. antibodies by Western Blot. Hematology evidenced light anemia, leukocytosis with eosinophilia, and lymphocytosis; biochemistry and enzyme activity revealed hypoalbuminemia with decreased albumin\/globulin ratio and elevated alkaline phosphatase values. Parasitic DNA extracted from skin biopsies of both donkeys demonstrated a homology of 100% with <jats:italic>Besnoitia<\/jats:italic> spp. This first clinical case of besnoitiosis in two donkeys in Italy both confirms the circulation of <jats:italic>Besnoitia<\/jats:italic> spp. in Italian equids and demonstrates that the distribution area of equine besnoitiosis in Europe could be wider than expected. Further studies are needed to infer its relevance, in relation to seroprevalence and clinical disease, and to identify the species of <jats:italic>Besnoitia<\/jats:italic> infecting donkeys. Besnoitiosis may be a neglected disease of donkeys in Europe: an early and accurate diagnosis is fundamental to implement adequate control measures to prevent a \u201csilent\u201d spread of <jats:italic>Besnoitia<\/jats:italic> spp. infection in equids populations.<\/jats:p>","DOI":"10.1007\/s00436-021-07089-9","type":"journal-article","created":{"date-parts":[[2021,3,16]],"date-time":"2021-03-16T03:03:23Z","timestamp":1615863803000},"page":"1811-1819","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Besnoitiosis in donkeys: an emerging parasitic disease of equids in Italy"],"prefix":"10.1007","volume":"120","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2013-3997","authenticated-orcid":false,"given":"Luca","family":"Villa","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alessia Libera","family":"Gazzonis","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Carlos","family":"Diezma-Diaz","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chiara","family":"Perlotti","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sergio Aurelio","family":"Zanzani","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Francesco","family":"Ferrucci","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gema","family":"\u00c1lvarez-Garc\u00eda","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Maria Teresa","family":"Manfredi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2021,3,16]]},"reference":[{"key":"7089_CR1","doi-asserted-by":"publisher","first-page":"407","DOI":"10.1016\/j.pt.2013.06.002","volume":"29","author":"G Alvarez-Garcia","year":"2013","unstructured":"Alvarez-Garcia G, Frey CF, Ortega-Mora LM, Schares G (2013) A century of bovine besnoitiosis: an unknown disease re-emerging in Europe. 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All procedures for the collection of biological specimens from live animals were accomplished following good clinical practices in the respect of animal welfare according to current legislation. The study was conducted with the approval of the Institutional Animal Care and Use Committee of Universit\u00e0 degli Studi di Milano (Prot. no OPBA_34_2017).","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethics approval"}},{"value":"Not applicable.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Consent to participate"}},{"value":"Not applicable.","order":4,"name":"Ethics","group":{"name":"EthicsHeading","label":"Consent for publication"}},{"value":"The authors declare no competing interests.","order":5,"name":"Ethics","group":{"name":"EthicsHeading","label":"Conflict of interest"}},{"value":"Informed consent was obtained from the owner of the animals.","order":6,"name":"Ethics","group":{"name":"EthicsHeading","label":"Informed consent"}}]}}