Molecular Characterization and Comparative Genomics of Two Staphylococcus pseudintermedius Strains from Humans in Egypt
Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Bacterial Strains, Media, and Growth Conditions
2.2. DNA Extraction, Library Preparation, and Whole-Genome Sequencing
2.3. Multi-Locus Sequence Typing
2.4. Pangenome Analysis
2.5. Antimicrobial Resistance Genes
3. Results
3.1. Genomic Features of S. pseudintermedius EGH1 and EGH2
3.2. Pangenome Analysis
3.3. Antimicrobial Resistance Genes
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
- Arefin, K.S.A.; Rana, E.A.; Islam, M.S.; Hossain, B.; Rahman, M.A.; Barua, H. Molecular Prevalence and Antimicrobial Resistance Profile of Staphylococcus aureus and Staphylococcus pseudintermedius Isolated From Hospital-Visited Cats. Vet. Med. Int. 2025, 2025, 4879266. [Google Scholar] [CrossRef] [PubMed]
- Dewulf, S.; Boyen, F.; Paepe, D.; Clercx, C.; Tilman, N.; Dewulf, J.; Boland, C. Antimicrobial Resistance Characterization of Methicillin-Resistant Staphylococcus aureus and Staphylococcus pseudintermedius Isolates from Clinical Cases in Dogs and Cats in Belgium. Antibiotics 2025, 14, 631. [Google Scholar] [CrossRef] [PubMed]
- Popa, I.; Iancu, I.; Iorgoni, V.; Degi, J.; Gligor, A.; Imre, K.; Tirziu, E.; Bochis, T.; Pop, C.; Plotuna, A.M.; et al. Antimicrobial Resistance Profile of Staphylococcus pseudintermedius Isolated from Dogs with Otitis Externa and Healthy Dogs: Veterinary and Zoonotic Implications. Antibiotics 2025, 14, 1027. [Google Scholar] [CrossRef] [PubMed]
- Borjesson, S.; Gomez-Sanz, E.; Ekstrom, K.; Torres, C.; Gronlund, U. Staphylococcus pseudintermedius can be misdiagnosed as Staphylococcus aureus in humans with dog bite wounds. Eur. J. Clin. Microbiol. Infect. Dis. 2015, 34, 839–844. [Google Scholar] [CrossRef]
- Yarbrough, M.L.; Lainhart, W.; Burnham, C.A. Epidemiology, Clinical Characteristics, and Antimicrobial Susceptibility Profiles of Human Clinical Isolates of Staphylococcus intermedius Group. J. Clin. Microbiol. 2018, 56, e01788-17. [Google Scholar] [CrossRef]
- Carroll, K.C.; Burnham, C.D.; Westblade, L.F. From canines to humans: Clinical importance of Staphylococcus pseudintermedius. PLoS Pathog. 2021, 17, e1009961. [Google Scholar] [CrossRef]
- Hanselman, B.A.; Kruth, S.A.; Rousseau, J.; Weese, J.S. Coagulase positive staphylococcal colonization of humans and their household pets. Can. Vet. J. 2009, 50, 954–958. [Google Scholar]
- Paul, N.C.; Moodley, A.; Ghibaudo, G.; Guardabassi, L. Carriage of methicillin-resistant Staphylococcus pseudintermedius in small animal veterinarians: Indirect evidence of zoonotic transmission. Zoonoses Public Health 2011, 58, 533–539. [Google Scholar] [CrossRef]
- Guardabassi, L.; Loeber, M.E.; Jacobson, A. Transmission of multiple antimicrobial-resistant Staphylococcus intermedius between dogs affected by deep pyoderma and their owners. Vet. Microbiol. 2004, 98, 23–27. [Google Scholar] [CrossRef]
- Peacock, S.J.; Paterson, G.K. Mechanisms of Methicillin Resistance in Staphylococcus aureus. Annu. Rev. Biochem. 2015, 84, 577–601. [Google Scholar] [CrossRef]
- Zehr, J.D.; Sun, Q.; Ceres, K.; Merrill, A.; Tyson, G.H.; Ceric, O.; Guag, J.; Pauley, S.; McQueary, H.C.; Sams, K.; et al. Population and pan-genomic analyses of Staphylococcus pseudintermedius identify geographic distinctions in accessory gene content and novel loci associated with AMR. Appl. Environ. Microbiol. 2025, 91, e0001025. [Google Scholar] [CrossRef] [PubMed]
- Abouelkhair, M.A.; Bemis, D.A.; Giannone, R.J.; Frank, L.A.; Kania, S.A. Characterization of a leukocidin identified in Staphylococcus pseudintermedius. PLoS ONE 2018, 13, e0204450. [Google Scholar] [CrossRef] [PubMed]
- Abouelkhair, M.A.; Bemis, D.A.; Giannone, R.J.; Frank, L.A.; Kania, S.A. Identification, cloning and characterization of SpEX exotoxin produced by Staphylococcus pseudintermedius. PLoS ONE 2019, 14, e0220301. [Google Scholar] [CrossRef] [PubMed]
- Ferrer, L.; Garcia-Fonticoba, R.; Perez, D.; Vines, J.; Fabregas, N.; Madronero, S.; Meroni, G.; Martino, P.A.; Martinez, S.; Mate, M.L.; et al. Whole genome sequencing and de novo assembly of Staphylococcus pseudintermedius: A pangenome approach to unravelling pathogenesis of canine pyoderma. Vet. Dermatol. 2021, 32, 654–663. [Google Scholar] [CrossRef]
- Saengsawang, P.; Tanonkaew, R.; Kimseng, R.; Nissapatorn, V.; Wintachai, P.; Rodriguez-Ortega, M.J.; Mitsuwan, W. Whole Genome Sequence Analysis of Multidrug-Resistant Staphylococcus aureus and Staphylococcus pseudintermedius Isolated from Superficial Pyoderma in Dogs and Cats. Antibiotics 2025, 14, 643. [Google Scholar] [CrossRef]
- Phophi, L.; Abouelkhair, M.; Jones, R.; Henton, M.; Qekwana, D.N.; Kania, S.A. The molecular epidemiology and antimicrobial resistance of Staphylococcus pseudintermedius canine clinical isolates submitted to a veterinary diagnostic laboratory in South Africa. PLoS ONE 2023, 18, e0290645. [Google Scholar] [CrossRef]
- Phophi, L.; Abouelkhair, M.A.; Jones, R.; Zehr, J.; Kania, S.A. Temporal changes in antibiotic resistance and population structure of methicillin-resistant Staphylococcus pseudintermedius between 2010 and 2021 in the United States. Comp. Immunol. Microbiol. Infect. Dis. 2023, 100, 102028. [Google Scholar] [CrossRef]
- Elaadli, H.; Badr, Y.; Raouf, M.; Kania, S.A.; Elsakhawy, O.K.; Altaib, H.; Abouelkhair, M.A. Isolation and Molecular Characterization of Three Staphylococcus pseudintermedius Strains from Dogs and Humans in Egypt. Curr. Microbiol. 2025, 82, 493. [Google Scholar] [CrossRef]
- Kearse, M.; Moir, R.; Wilson, A.; Stones-Havas, S.; Cheung, M.; Sturrock, S.; Buxton, S.; Cooper, A.; Markowitz, S.; Duran, C.; et al. Geneious Basic: An integrated and extendable desktop software platform for the organization and analysis of sequence data. Bioinformatics 2012, 28, 1647–1649. [Google Scholar] [CrossRef]
- Gurevich, A.; Saveliev, V.; Vyahhi, N.; Tesler, G. QUAST: Quality assessment tool for genome assemblies. Bioinformatics 2013, 29, 1072–1075. [Google Scholar] [CrossRef]
- Merkel, D. Docker: Lightweight Linux containers for consistent development and deployment. Linux J. 2014, 2014, 2. [Google Scholar]
- Jolley, K.A.; Bray, J.E.; Maiden, M.C.J. Open-access bacterial population genomics: BIGSdb software, the PubMLST.org website and their applications. Wellcome Open Res. 2018, 3, 124. [Google Scholar] [CrossRef] [PubMed]
- Seemann, T. Prokka: Rapid prokaryotic genome annotation. Bioinformatics 2014, 30, 2068–2069. [Google Scholar] [CrossRef]
- Page, A.J.; Cummins, C.A.; Hunt, M.; Wong, V.K.; Reuter, S.; Holden, M.T.; Fookes, M.; Falush, D.; Keane, J.A.; Parkhill, J. Roary: Rapid large-scale prokaryote pan genome analysis. Bioinformatics 2015, 31, 3691–3693. [Google Scholar] [CrossRef] [PubMed]
- Feldgarden, M.; Brover, V.; Gonzalez-Escalona, N.; Frye, J.G.; Haendiges, J.; Haft, D.H.; Hoffmann, M.; Pettengill, J.B.; Prasad, A.B.; Tillman, G.E.; et al. AMRFinderPlus and the Reference Gene Catalog facilitate examination of the genomic links among antimicrobial resistance, stress response, and virulence. Sci. Rep. 2021, 11, 12728. [Google Scholar] [CrossRef]
- Gomez-Sanz, E.; Torres, C.; Lozano, C.; Zarazaga, M. High diversity of Staphylococcus aureus and Staphylococcus pseudintermedius lineages and toxigenic traits in healthy pet-owning household members. Underestimating normal household contact? Comp. Immunol. Microbiol. Infect. Dis. 2013, 36, 83–94. [Google Scholar] [CrossRef]
- Kronbichler, A.; Blane, B.; Holmes, M.A.; Wagner, J.; Parkhill, J.; Peacock, S.J.; Jayne, D.R.W.; Harrison, E.M. Nasal carriage of Staphylococcus pseudintermedius in patients with granulomatosis with polyangiitis. Rheumatology 2019, 58, 548–550. [Google Scholar] [CrossRef]
- Kuan, E.C.; Yoon, A.J.; Vijayan, T.; Humphries, R.M.; Suh, J.D. Canine Staphylococcus pseudintermedius sinonasal infection in human hosts. Int. Forum Allergy Rhinol. 2016, 6, 710–715. [Google Scholar] [CrossRef]
- Alhadz, G.G.; Salasia, S.I.O.; Lestari, F.B.; Yosyana, A.R.P.; Wasissa, M.; Setianingrum, Y.R.; Widayanti, R. Occurrence, molecular confirmation, and multidrug resistance of methicillin-resistant Staphylococcus aureus and Staphylococcus pseudintermedius in companion animals in Indonesia. Vet. World 2026, 19, 324–338. [Google Scholar] [CrossRef]
- Afema, J.A.; Mastromonaco, C.; Davis, M.A.; Stone, D.M.; Jones, L.P.; Paterson, T.E.; Perea, M.L.; Butler, B.P. Emergence of methicillin resistant Staphylococcus pseudintermedius in dogs sampled in 2018 in the island nation of Grenada, West Indies. Front. Vet. Sci. 2026, 13, 1761713. [Google Scholar] [CrossRef]
- Douan, J.; Kohler, C.; Haralambiev, L.; Idelevich, E.A.; Becker, K. Comparative phylogenetic, antimicrobial resistance, and clinical characterization of human spondylodiscitis-associated Staphylococcus pseudintermedius. Front. Microbiol. 2026, 17, 1735075. [Google Scholar] [CrossRef]
- Wiedenbeck, J.; Cohan, F.M. Origins of bacterial diversity through horizontal genetic transfer and adaptation to new ecological niches. FEMS Microbiol. Rev. 2011, 35, 957–976. [Google Scholar] [CrossRef]
- Di Carluccio, C.; Soriano-Maldonado, P.; Berni, F.; de Haas, C.J.C.; Temming, A.R.; Hendriks, A.; Ali, S.; Molinaro, A.; Silipo, A.; van Sorge, N.M.; et al. Antibody Recognition of Different Staphylococcus aureus Wall Teichoic Acid Glycoforms. ACS Cent. Sci. 2022, 8, 1383–1392. [Google Scholar] [CrossRef]
- Atilano, M.L.; Yates, J.; Glittenberg, M.; Filipe, S.R.; Ligoxygakis, P. Wall teichoic acids of Staphylococcus aureus limit recognition by the drosophila peptidoglycan recognition protein-SA to promote pathogenicity. PLoS Pathog. 2011, 7, e1002421. [Google Scholar] [CrossRef]



| EGH1 | EGH2 | |
|---|---|---|
| Number of reads | 9,499,989 | 9,567,531 |
| Genome length | 2,536,812 bp | 2,636,008 bp |
| N50 | 141,100 bp | 139,200 bp |
| Number of contigs | 41 | 99 |
| GC percent | 37.5% | 37.5% |
| Genome coverage | 9.22× | 9.29× |
| Genes | 2424 | 2596 |
| Isolate | ack | cpn60 | fdh | pta | pura | Sar | tuf | Sequence Type (ST) |
|---|---|---|---|---|---|---|---|---|
| S. pseudintermdius EGH1 | 6 | 2 | 1 | 1 | 107 | 1 | 2 | 3037 |
| S. pseudintermdius EGH2 | 1 | 18 | 2 | 2 | 3 | 1 | 1 | 2874 |
| Antimicrobial Class | Gene | EGH1 | EGH2 |
|---|---|---|---|
| Aminoglycoside | ant(6)-Ia | + | − |
| aph(3′)-IIIa | + | − | |
| β-Lactam | blaI | + | + |
| blaPC1 | + | + | |
| blaR1 | + | + | |
| Chloramphenicol | catA | + | − |
| Fusidic acid | fusC | + | + |
| Macrolide–lincosamide–streptogramin B | erm(B) | + | − |
| Streptothricin | sat4 | + | − |
| Tetracycline | tet(M) | + | − |
| tet(K) | − | + | |
| Trimethoprim | dfrG | − | + |
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Elsakhawy, O.K.; Elaadli, H.; Badr, Y.; Raouf, M.; Kania, S.A.; Altaib, H.; Abouelkhair, M.A. Molecular Characterization and Comparative Genomics of Two Staphylococcus pseudintermedius Strains from Humans in Egypt. Vet. Sci. 2026, 13, 424. https://doi.org/10.3390/vetsci13050424
Elsakhawy OK, Elaadli H, Badr Y, Raouf M, Kania SA, Altaib H, Abouelkhair MA. Molecular Characterization and Comparative Genomics of Two Staphylococcus pseudintermedius Strains from Humans in Egypt. Veterinary Sciences. 2026; 13(5):424. https://doi.org/10.3390/vetsci13050424
Chicago/Turabian StyleElsakhawy, Ola K., Haitham Elaadli, Yassien Badr, May Raouf, Stephen A. Kania, Hend Altaib, and Mohamed A. Abouelkhair. 2026. "Molecular Characterization and Comparative Genomics of Two Staphylococcus pseudintermedius Strains from Humans in Egypt" Veterinary Sciences 13, no. 5: 424. https://doi.org/10.3390/vetsci13050424
APA StyleElsakhawy, O. K., Elaadli, H., Badr, Y., Raouf, M., Kania, S. A., Altaib, H., & Abouelkhair, M. A. (2026). Molecular Characterization and Comparative Genomics of Two Staphylococcus pseudintermedius Strains from Humans in Egypt. Veterinary Sciences, 13(5), 424. https://doi.org/10.3390/vetsci13050424

