Mechanisms of Linezolid Resistance Among Enterococci of Clinical Origin in Spain—Detection of optrA- and cfr(D)-Carrying E. faecalis
Abstract
:1. Introduction
2. Materials and Methods
2.1. Bacterial Collection
2.2. Antimicrobial Resistance Phenotype
2.3. Mechanisms of Linezolid Resistance
2.4. Molecular Characterization
2.5. Clonal Relatedness
2.6. Conjugation Assays
3. Results
3.1. Mechanisms of Oxazolidinone Resistance
3.2. Molecular Characterization and Clonal Relatedness
3.3. Resistance to Non-Oxazolidinone Agents
3.4. Virulence Profile
3.5. Genetic Environment of the optrA and cfr(D) Genes
3.6. Transferability of the optrA and cfr(D) Genes
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Isolate | Hospital 1 | Year | Type of Sample | MLST/CC | Linezolid MIC (mg/L) | Mechanisms of Linezolid Resistance 2 | Antimicrobial Resistance Phenotype 3 | Antimicrobial Resistance Genotype | Virulence Genes |
---|---|---|---|---|---|---|---|---|---|
C9884 | HSJ | 2017 | Urine | ST585/CC4 | 16 | optrA | ERY-CLI-GEN-KAN-STR-TET-CHL-FFC-LZD-TDZ-SXT-CIP 4-LEV | erm(B), lnu(B), lsa(A), lsa(E), aac(6′)-Ie-aph(2″)-Ia, aph(3″)-III, ant(6)-Ia, str, tet(M), tet(L), optrA, fexA, dfrG | gelE, ace, agg, efaAfs, elrA, srtA, ccf, cob, cad, camE, ebpA, ebpC, fsrB, tpx |
C8946 | HUG | 2017 | Leg wound exudate | ST480 | 16 | optrA | ERY-CLI-GEN-KAN-STR-TET-CHL-FFC-LZD-TDZ-SXT-CIP 4,5-LEV | erm(B), lnu(B), lsa(A), lsa(E), aac(6′)-Ie-aph(2″)-Ia, aph(3″)-III, ant(6)-Ia, tet(M), tet(L), optrA, fexA, dfrG | hylA, ace, efaAfs, elrA, srtA, ccf, cob, cad, camE, ebpA, ebpB, tpx |
C9901 | HUG | 2017 | Urine | ST21/CC21 | 16 | optrA | ERY-CLI-KAN-STR-TET-CHL-FFC-LZD-TDZ | erm(B), lnu(B), lsa(A), lsa(E), aph(3″)-III, ant(6)-Ia, tet(M), tet(L), optrA, fexA | gelE, hylA, hylB, ace, agg, cylL, efaAfs, elrA, srtA, ccf, cob, cad, camE, ebpA, ebpC, fsrB, tpx |
C9951 | HSP | 2017 | Urine | ST16 | 8 | optrA (Y176D, T481P); 23S rDNA (G2576T) | ERY-CLI-GEN-KAN-STR-TET-CHL-FFC-LZD-TDZ-SXT | erm(A)-like, erm(B), lnu(B), lsa(A), lsa(E), aac(6′)-Ie-aph(2″)-Ia, aph(3″)-III, ant(6)-Ia, tet(M), tet(L), tet(O), optrA, fexA, dfrG | ace, agg, cylA, efaAfs, elrA, srtA, ccf, cob, cad, camE, ebpA, ebpC, tpx |
C9952 | HSP | 2017 | Urine | ST35 | 8 | optrA (Y176D, T481P) | ERY-CLI-GEN-KAN-STR-TET CHL-FFC-LZD-TDZ-CIP 4,6-LEV | erm(A)-like, erm(B), lnu(B), lsa(A), lsa(E), aac(6′)-Ie-aph(2″)-Ia, aph(3″)-III, ant(6)-Ia, tet(M), tet(L), optrA, fexA | gelE, hylB, ace, cylA, cylL, cylM, efaAfs, elrA, srtA, ccf, cob, cad, camE, ebpA, ebpB, fsrB, tpx |
C9736 | HVC | 2017 | Urine | ST585/CC4 | 16 | optrA | ERY-CLI-GEN-KAN-STR-TET-CHL-FFC-LZD-TDZ-SXT-CIP 4-LEV | erm(B), lnu(B), lsa(A), lsa(E), aac(6′)-Ie-aph(2″)-Ia, aph(3″)-III, ant(6)-Ia, str, tet(M), tet(L), optrA, fexA, dfrG | gelE, ace, agg, cylL, efaAfs, elrA, srtA, ccf, cob, cad, camE, ebpA, ebpC, fsrB, tpx |
C9737 | HVC | 2017 | Urine | ST896 | 16 | optrA | ERY-CLI-KAN-STR-TET-CHL-FFC-LZD-TDZ-CIP 4,5-LEV | erm(B), lnu(B), lsa(A), lsa(E), aph(3″)-III, ant(6)-Ia, tet(M), optrA, fexA, catpC221 | esp, gelE, hylA, hylB, ace, agg, efaAfs, elrA, srtA, ccf, cob, cad, camE, ebpA, ebpB, tpx |
X523 | HVC | 2018 | Otic exudate | ST480 | 8 | optrA (T572P) | ERY-CLI-TET-CHL-FFC-LZD-TDZ-SXT-CIP 4,5-LEV | lsa(A), lsa(E), tet(M), tet(L), optrA, fexA, dfrG | hylA, ace, efaAfs, elrA, srtA, ccf, cob, cad, camE, ebpA, ebpB, tpx |
X524 | HVC | 2018 | Urine | ST480 | 8 | optrA | ERY-CLI-GEN-KAN-STR-TET-CHL-FFC-LZD-TDZ-SXT-CIP 4,5-LEV | erm(B), lsa(A), lsa(E), aac(6′)-Ie-aph(2″)-Ia, aph(3″)-III, ant(6)-Ia, tet(M), tet(L), optrA, fexA, dfrG | hylA, ace, agg, cylL, cylM, efaAfs, elrA, srtA, ccf, cob, cad, camE, ebpA, ebpB, tpx |
X525 | HVC | 2018 | Ulcer | ST585/CC4 | 8 | optrA | ERY-CLI-GEN-KAN-STR-TET-CHL-FFC-LZD-TDZ-SXT-CIP 4-LEV | erm(B), lnu(B), lsa(A), lsa(E), aac(6′)-Ie-aph(2″)-Ia, aph(3″)-III, ant(6)-Ia, str, tet(M), tet(L), optrA, fexA, catA, dfrG | gelE, ace, agg, cylL, efaAfs, elrA, srtA, cob, cad, camE, ebpA, ebpC, fsrB, tpx |
X526 | HVC | 2018 | Urine | ST896 | 4 | optrA | ERY-CLI-KAN-STR-TET-CHL-FFC-LZD-TDZ-CIP 4,5-LEV | erm(B), lnu(B), lsa(A), lsa(E), aph(3″)-III, ant(6)-Ia, tet(M), optrA, fexA, catpC221 | gelE, hylA, hylB, ace, cylA, cylL, efaAfs, elrA, srtA, ccf, cob, cad, camE, ebpA, ebpB, fsrB, tpx |
X527 | HVC | 2018 | Ulcer | ST6/CC2 | 16 | optrA | ERY-CLI-GEN-KAN-STR-TET-CHL-FFC-LZD-TDZ-CIP 4,5-LEV | erm(B), lsa(A), aac(6′)-aph(2″), aph(3″)-III, ant(6)-Ia, tet(M), optrA, fexA | gelE, hylA, hylB, ace, agg, efaAfs, elrA, srtA, cob, cad, camE, ebpA, ebpB, ebpC, fsrB, tpx |
X528 | HVC | 2018 | Abscess | ST480 | 16 | optrA; cfr(D) | ERY-CLI-KAN-STR-TET-CHL-FFC-LZD-TDZ-SXT-CIP 4,5-LEV | erm(B), lsa(A), aph(3′’)-III, ant(6)-Ia, tet(M), tet(L), cfr(D), optrA, fexA, dfrG | hylA, ace, agg, efaAfs, elrA, srtA, ccf, cob, cad, camE, ebpA, ebpB, tpx |
Isolate | Hospital 1 | Year | Type of Sample | PFGE Pattern | MLST | LZD MIC (mg/L) | Mechanisms of LZD Resistance 2 | Antimicrobial Resistance Phenotype 3 | Antimicrobial Resistance Genotype 4 | Virulence Genes |
---|---|---|---|---|---|---|---|---|---|---|
C9902 | HLB | 2017 | Abscess | A | ST17 | 8 | 23S rDNA (G2576T) | PEN 5-AMP-ERY-CLI-KAN-STR-CIP 8-LEV-LZD-SXT | erm(B), msrC, aac(6′)-Ii, aph(3″)-III, ant(6)-Ia, dfrG | esp, hyl |
C9903 | HLB | 2017 | Surgical wound | B | ST262 | 16 | 23S rDNA (G2576T) | PEN 5-AMP-ERY-CLI-GEN-KAN-STR-TET-CIP 8-LEV-LZD-SXT | msr(C), lsa(E), aac(6′)-Ie-aph(2″)-Ia, aac(6′)-Ii, ant(6)-Ia, tet(M), tet(L), dfrG | |
C10262 | HLB | 2017 | Blood | C | ST117 | 32 | 23S rDNA (G2576T) | PEN 6-AMP-ERY-CLI-KAN-STR-CIP 8-LEV-LZD-SXT | erm(B), msr(C), aac(6′)-Ii, aph(3″)-III, ant(6)-Ia, dfrG | esp, hyl |
C9950 | HSP | 2017 | Blood | D | ST117 | 4 | 23S rDNA (G2576T) | PEN 7-AMP-ERY-CLI-KAN-STR-CIP 8-LEV-LZD-SXT | erm(B), msr(C), aac(6′)-Ii, aph(3″)-III, ant(6)-Ia, dfrG | esp, hyl |
C9953 | HSP | 2017 | Urine | E | ST117 | 16 | 23S rDNA (G2576T) | PEN I,5-AMP-ERY-CLI-KAN-STR-CIP 8-LEV-LZD-SXT | erm(B), msr(C), aac(6′)-Ii, aph(3″)-III, ant(6)-Ia, dfrG | hyl |
C10269 | HRV | 2017 | Surgical wound | F | ST117 | 16 | 23S rDNA (G2576T) | PEN 6-AMP-ERY-CLI-KAN-STR-CIP 8-LEV-LZD-SXT | erm(B), msr(C), aac(6′)-Ii, aph(3″)-III, ant(6)-Ia, dfrG | esp, hyl |
Strain | Genotype | MIC (mg/L) 1 | ||||||
---|---|---|---|---|---|---|---|---|
ERY | CLI | TET | CHL | FFC | LZD | TDZ | ||
E. faecalisJH2-2 (recipient) | - | 1 | 2 | 0.125 | 4 | 4 | 2 | 1 |
E. faecalisC9884 | erm(B), lnu(B), lsa(A), lsa(E), aac(6′)-aph(2″), aph(3″)-III, ant(6)-Ia, str, tet(M), tet(L), optrA, fexA, dfrG | >256 | 256 | 128 | 64 | 64 | 16 | 1 |
E. faecalisTC-C9884 | optrA, fexA | 1 | 2 | 0.125 | 32 | 64 | 8 | 1 |
E. faecalisC9901 | erm(B), lnu(B), lsa(A), lsa(E), aph(3″)-III, ant(6)-Ia, tet(M), tet(L), optrA, fexA | >256 | >256 | >256 | 32 | 64 | 16 | 1 |
E. faecalisTC-C9901 | optrA, fexA | 1 | 2 | 0.125 | 32 | 64 | 4 | 1 |
E. faecalisC9951 | erm(A)-like, erm(B), lnu(B), lsa(A), lsa(E), aac(6′)-aph(2″), aph(3″)-III, ant(6)-Ia, tet(M), tet(L), tet(O), optrA, fexA, dfrG | >256 | >256 | 128 | 64 | 64 | 8 | 1 |
E. faecalisTC-C9951 | erm(A)-like, tet(L), tet(O), optrA, fexA | 256 | >256 | 128 | 64 | 64 | 4 | 1 |
E. faecalisC9952 | erm(A)-like, erm(B), lnu(B), lsa(A), lsa(E), aac(6′)-aph(2″), aph(3″)-III, ant(6)-Ia, tet(M), tet(L), optrA, fexA | 256 | >256 | >256 | 32 | 64 | 8 | 1 |
E. faecalisTC-C9952 | erm(A)-like, optrA, fexA | 128 | >256 | 0.125 | 32 | 64 | 4 | 1 |
E. faecalisC9736 | erm(B), lnu(B), lsa(A), lsa(E), aac(6′)-aph(2″), aph(3″)-III, ant(6)-Ia, str, tet(M), tet(L), optrA, fexA, dfrG | >256 | 128 | 256 | 128 | 32 | 16 | 1 |
E. faecalisTC-C9736 | optrA, fexA | 1 | 2 | 0.125 | 64 | 32 | 8 | 1 |
E. faecalisC9737 | erm(B), lnu(B), lsa(A), lsa(E), aph(3″)-III, ant(6)-Ia, tet(M), optrA, fexA, catpC221 | >256 | 128 | 128 | 32 | 64 | 16 | 1 |
E. faecalisTC-C9737 | optrA, fexA | 1 | 2 | 0.125 | 32 | 32 | 8 | 1 |
E. faecalisX523 | lsa(A), lsa(E), tet(M), tet(L), optrA, fexA, dfrG | 128 | >256 | 128 | 32 | 64 | 8 | 1 |
E. faecalisTC-X523 | optrA, fexA | 1 | 2 | 0.125 | 32 | 64 | 8 | 1 |
E. faecalisX524 | erm(B), lsa(A), lsa(E), aac(6′)-aph(2″), aph(3″)-III, ant(6)-Ia, tet(M), tet(L), optrA, fexA, dfrG | 256 | 256 | 256 | 32 | 64 | 8 | 1 |
E. faecalisTC-X524 | optrA, fexA | 1 | 2 | 0.125 | 32 | 64 | 8 | 1 |
E. faecalisX525 | erm(B), lnu(B), lsa(A), lsa(E), aac(6′)-aph(2″), aph(3″)-III, ant(6)-Ia, str, tet(M), tet(L), optrA, fexA, catA, dfrG | >256 | 128 | 128 | 64 | 64 | 8 | 1 |
E. faecalisTC-X525 | optrA, fexA | 1 | 2 | 0.125 | 32 | 32 | 8 | 1 |
E. faecalisX526 | erm(B), lnu(B), lsa(A), lsa(E), aph(3″)-III, ant(6)-Ia, tet(M), optrA, fexA, catpC221 | >256 | 128 | 128 | 32 | 64 | 4 | 1 |
E. faecalisTC-X526 | optrA, fexA | 1 | 2 | 0.125 | 32 | 64 | 4 | 1 |
E. faecalisX527 | erm(B), lsa(A), aac(6′)-aph(2″), aph(3″)-III, ant(6)-Ia, tet(M), optrA, fexA | >256 | 256 | 128 | 32 | 64 | 16 | 1 |
E. faecalisTC-X527 | optrA, fexA | 1 | 2 | 0.125 | 32 | 64 | 8 | 1 |
E. faecalisX528 | erm(B), lsa(A), aph(3″)-III, ant(6)-Ia, tet(M), tet(L), cfr(D), optrA, fexA, dfrG | >256 | >256 | >256 | 32 | 64 | 16 | 1 |
E. faecalisTC-X528 | optrA, fexA | 1 | 2 | 0.125 | 32 | 64 | 8 | 1 |
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Ruiz-Ripa, L.; Feßler, A.T.; Hanke, D.; Eichhorn, I.; Azcona-Gutiérrez, J.M.; Pérez-Moreno, M.O.; Seral, C.; Aspiroz, C.; Alonso, C.A.; Torres, L.; et al. Mechanisms of Linezolid Resistance Among Enterococci of Clinical Origin in Spain—Detection of optrA- and cfr(D)-Carrying E. faecalis. Microorganisms 2020, 8, 1155. https://doi.org/10.3390/microorganisms8081155
Ruiz-Ripa L, Feßler AT, Hanke D, Eichhorn I, Azcona-Gutiérrez JM, Pérez-Moreno MO, Seral C, Aspiroz C, Alonso CA, Torres L, et al. Mechanisms of Linezolid Resistance Among Enterococci of Clinical Origin in Spain—Detection of optrA- and cfr(D)-Carrying E. faecalis. Microorganisms. 2020; 8(8):1155. https://doi.org/10.3390/microorganisms8081155
Chicago/Turabian StyleRuiz-Ripa, Laura, Andrea T. Feßler, Dennis Hanke, Inga Eichhorn, José Manuel Azcona-Gutiérrez, Mar Olga Pérez-Moreno, Cristina Seral, Carmen Aspiroz, Carla Andrea Alonso, Luis Torres, and et al. 2020. "Mechanisms of Linezolid Resistance Among Enterococci of Clinical Origin in Spain—Detection of optrA- and cfr(D)-Carrying E. faecalis" Microorganisms 8, no. 8: 1155. https://doi.org/10.3390/microorganisms8081155