Molecular Characterization and Comparative Genomics of Two Staphylococcus pseudintermedius Strains from Humans in Egypt

Staphylococcus pseudintermedius is an opportunistic bacterium previously associated with dogs but has recently been found in human infections, raising zoonotic concerns. Genomic characterization of human S. pseudintermedius isolates can provide preliminary information on antibiotic resistance, pathogenicity, and genomic features relevant to host range. Two S. pseudintermedius isolates (hereafter referred to as S. pseudintermedius EGH1 and S. pseudintermedius EGH2) from human clinical samples in Egypt were sequenced using the Illumina NovaSeq X Plus platform. To assess genetic relatedness to human S. pseudintermedius isolates worldwide, multilocus sequence typing (MLST), pangenome analysis, and antimicrobial resistance gene profiling were performed. The sequencing produced a total of 9,499,989 reads for S. pseudintermedius EGH1 and 9,567,531 reads for S. pseudintermedius EGH2. Sequences were assembled with Geneious Prime® 2025 and annotated using NCBI Prokaryotic Genome Annotation Pipeline v6.10. Pangenome analysis identified 9574 genes, comprising 1681 core genes (17.56%), 180 soft-core genes (1.88%), 837 shell genes (8.74%), and 6876 cloud genes (71.82%). MLST was conducted on human S. pseudintermedius genome assemblies using MLST v2.23.0. The analysis revealed both isolates as novel sequence types: S. pseudintermedius EGH1 was assigned ST-3037 with a new allele (purA-107), and S. pseudintermedius EGH2 was assigned ST-2874. Clonal relationships among S. pseudintermedius isolates were evaluated using the eBURST algorithm. This study presents the first next-generation genome sequencing and comparative genomic analysis of S. pseudintermedius isolates from humans in Egypt. Future studies integrating genomic, epidemiological, and phenotypic data are required.