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Synchrotron-Radiation-Based Fourier Transform Infrared Microspectroscopy as a Tool for the Differentiation between Staphylococcal Small Colony Variants

ORCID
0000-0002-6299-5301
Affiliation
Department of Pharmaceutics and Pharmaceutical Technology, School of Pharmacy, The University of Jordan, Amman 11942, Jordan
Al-Bakri, Amal G.;
ORCID
0000-0002-4688-7052
Affiliation
Department of Pharmaceutical Sciences, School of Pharmacy, The University of Jordan, Amman 11942, Jordan
Dahabiyeh, Lina A.;
ORCID
0000-0002-5878-443X
Affiliation
Department of Pharmaceutics and Pharmaceutical Technology, School of Pharmacy, The University of Jordan, Amman 11942, Jordan
Khalil, Enam;
ORCID
0000-0002-4691-0427
Affiliation
Department of Pharmaceutics and Pharmaceutical Technology, School of Pharmacy, The University of Jordan, Amman 11942, Jordan
Jaber, Deema;
ORCID
0000-0003-0854-1920
Affiliation
SESAME Synchrotron (Synchrotron-Light for Experimental Science and Applications in the Middle East), Allan 19252, Jordan
Kamel, Gihan;
Affiliation
Institute of Medical Microbiology, University Hospital Münster, 48149 Münster, Germany
Schleimer, Nina;
Affiliation
Friedrich Loeffler-Institute of Medical Microbiology, University Medicine Greifswald, 17475 Greifswald, Germany
Kohler, Christian;
ORCID
0000-0002-6391-1341
Affiliation
Institute of Medical Microbiology, University Hospital Münster, 48149 Münster, Germany
Becker, Karsten

Small colony variants (SCVs) are clinically significant and linked to persistent infections. In this study, synchrotron-radiation-based Fourier transform infrared (SR-FTIR) is used to investigate the microspectroscopic differences between the SCVs of Staphylococcus aureus ( S. aureus ) and diabetic foot Staphylococcus epidermidis ( S. epidermidis ) in two main IR spectral regions: (3050–2800 cm −1 ), corresponding to the distribution of lipids, and (1855–1500 cm −1 ), corresponding to the distribution of protein amide I and amide II and carbonyl vibrations. SR-FTIR successfully discriminated between the two staphylococcal species and between the SCV and the non-SCV strains within the two IR spectral regions. Combined S. aureus SCVs (SCVhMu) showed a higher protein content relative to the non-SCV wild type. Complemented S. aureus SCV showed distinguishable differences from the SCVhMu and the wild type, including a higher content of unsaturated fatty acids. An increase in the CH 2 /CH 3 ratio was detected in S. epidermidis SCV samples compared to the standard control. Protein secondary structure in standard S. epidermidis and SCVs consisted mainly of an α-helix; however, a new shoulder at 1635 cm −1 , assigned to β-sheets, was evident in the SCV. In conclusion, SR-FTIR is a powerful method that can discriminate between staphylococci species and to differentiate between SCVs and their corresponding natural strains.

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