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Understanding its genetic and phenotypic properties, such as virulence factors and antimicrobial resistance (AMR) profiles, is critical for infection control and vaccine development. In this study, we isolated and molecularly characterized three <jats:italic>S. pseudintermedius<\/jats:italic> isolates from dogs (hereafter referred to as <jats:italic>S. pseudintermedius<\/jats:italic> D8) and humans (hereafter referred to as <jats:italic>S. pseudintermedius<\/jats:italic> H10 and <jats:italic>S. pseudintermedius<\/jats:italic> H11) in Egypt. All three isolates showed 100% sequence identity with the <jats:italic>nuc<\/jats:italic> gene of the <jats:italic>S. pseudintermedius<\/jats:italic> SP_11304-3A reference genome. Multilocus sequence typing (MLST) revealed novel sequence types (STs) in the three isolates. The AMR determinants varied substantially among the isolates. While the <jats:italic>mecA<\/jats:italic> gene was absent, <jats:italic>blaZ<\/jats:italic> was detected in the canine isolate, indicating beta-lactamase-mediated penicillin resistance. Additionally, <jats:italic>tetK<\/jats:italic> and <jats:italic>tetM<\/jats:italic> genes were found conferring tetracycline resistance in different isolates. Resistance genes for aminoglycosides, chloramphenicol, fusidic acid, macrolides, streptothricin, and trimethoprim were also identified. All isolates were positive for key virulence genes, including immune evasion (<jats:italic>AdsA<\/jats:italic>), coagulase (<jats:italic>coa<\/jats:italic>), immunoglobulin-binding protein (<jats:italic>sbi\/spsK<\/jats:italic>), exfoliative toxin (<jats:italic>speta<\/jats:italic>), enterotoxins (<jats:italic>se-int<\/jats:italic> and <jats:italic>siet<\/jats:italic>), fibrinogen binding protein gene (<jats:italic>fnbB<\/jats:italic>), and two-component pore-forming leukocidin genes (<jats:italic>lukF<\/jats:italic> and<jats:italic> lukS<\/jats:italic>). The <jats:italic>S. pseudintermedius<\/jats:italic> H11 isolate uniquely harbored the neuraminidase gene (<jats:italic>nanB<\/jats:italic>), while none of the isolates contained the gene coding for immunoglobulin G binding protein (<jats:italic>spsQ<\/jats:italic>). These findings highlight the differences in virulence and antimicrobial resistance genes among these <jats:italic>S. pseudintermedius<\/jats:italic> isolates, underlining the need for global surveillance and molecular characterization of this pathogen.<\/jats:p>","DOI":"10.1007\/s00284-025-04477-7","type":"journal-article","created":{"date-parts":[[2025,9,6]],"date-time":"2025-09-06T09:22:25Z","timestamp":1757150545000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Isolation and Molecular Characterization of Three Staphylococcus pseudintermedius Strains from Dogs and Humans in Egypt"],"prefix":"10.1007","volume":"82","author":[{"given":"Haitham","family":"Elaadli","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0615-0822","authenticated-orcid":false,"given":"Yassien","family":"Badr","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"May","family":"Raouf","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Stephen A.","family":"Kania","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ola K.","family":"Elsakhawy","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hend","family":"Altaib","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0109-6000","authenticated-orcid":false,"given":"Mohamed A.","family":"Abouelkhair","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2025,9,6]]},"reference":[{"issue":"3","key":"4477_CR1","doi-asserted-by":"publisher","first-page":"603","DOI":"10.1128\/CMR.00134-14","volume":"28","author":"SYC Tong","year":"2015","unstructured":"Tong SYC, Davis JS, Eichenberger E, Holland TL, Fowler VG (2015) Staphylococcus aureus infections: epidemiology, pathophysiology, clinical manifestations, and management. 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