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Characterization of OXA232-Producing Carbapenem-Resistant Klebsiella pneumoniae: Genomic Analysis and Virulence Assessment Cover

Characterization of OXA232-Producing Carbapenem-Resistant Klebsiella pneumoniae: Genomic Analysis and Virulence Assessment

Open Access
|Mar 2025

Figures & Tables

Fig. 1.

Circular representation of IncFII/FIB-type plasmids in ST231 isolates.
The plasmid pIncFIB-Q1 spans 127,300 base pairs (bp). The map is divided into several tracks, each representing different genomic features.
The outermost tracks mark specific genes with resistance markers (e.g., blaKPC, qnrB19), the middle tracks show genes with varying expression levels indicated by different color shades, and the innermost tracks depict GC content and GC skew. Each track uses a distinct color code, as the legend explains, to represent gene density, antibiotic resistance genes, virulence factors, and mobile genetic elements.
Circular representation of IncFII/FIB-type plasmids in ST231 isolates. The plasmid pIncFIB-Q1 spans 127,300 base pairs (bp). The map is divided into several tracks, each representing different genomic features. The outermost tracks mark specific genes with resistance markers (e.g., blaKPC, qnrB19), the middle tracks show genes with varying expression levels indicated by different color shades, and the innermost tracks depict GC content and GC skew. Each track uses a distinct color code, as the legend explains, to represent gene density, antibiotic resistance genes, virulence factors, and mobile genetic elements.

Fig. 2.

Phylogenetic analysis and genotypic characterization of bacterial strains.
The phylogenetic relationships and distribution of resistance genes and virulence factors across ten bacterial strains (Kp1 to Kp13) are displayed.
The phylogenetic tree (left) illustrates evolutionary distances based on whole-genome sequencing data, while the heatmap (right) indicates the presence (colored) and absence (white) of specific resistance genes (green) and virulence factors (blue).
Phylogenetic analysis and genotypic characterization of bacterial strains. The phylogenetic relationships and distribution of resistance genes and virulence factors across ten bacterial strains (Kp1 to Kp13) are displayed. The phylogenetic tree (left) illustrates evolutionary distances based on whole-genome sequencing data, while the heatmap (right) indicates the presence (colored) and absence (white) of specific resistance genes (green) and virulence factors (blue).

Fig. 3.

Phylogenetic analysis of global ST231 Klebsiella pneumoniae isolates.
The phylogenetic tree constructed from core SNP analyses of 264 global ST231 isolates is color-coded to represent various clades (R1: Clades), geographical origins (R2: Country and extra: Province), and specific genetic characteristics (R3: OXA-48-like types; R4: Yersiniabactin types; R5: Aerobactin type, R6: K locus, R7: O locus). Each concentric ring corresponds to different attributes, such as antibiotic resistance genes, toxin production genes, and other relevant genetic markers. The innermost circle shows phylogenetic relationships based on whole-genome sequencing data.
Phylogenetic analysis of global ST231 Klebsiella pneumoniae isolates. The phylogenetic tree constructed from core SNP analyses of 264 global ST231 isolates is color-coded to represent various clades (R1: Clades), geographical origins (R2: Country and extra: Province), and specific genetic characteristics (R3: OXA-48-like types; R4: Yersiniabactin types; R5: Aerobactin type, R6: K locus, R7: O locus). Each concentric ring corresponds to different attributes, such as antibiotic resistance genes, toxin production genes, and other relevant genetic markers. The innermost circle shows phylogenetic relationships based on whole-genome sequencing data.

Fig. 4.

Weight change and survival analysis of mice infected with ST231 Klebsiella pneumoniae.
Six-week-old male C57BL/6 mice, grouped in trios, were inoculated intranasally with 30 μl of 108 CFU/ml bacterial suspension. Health monitoring occurred daily for seven days.
a)Daily average weight fluctuations among mice infected with different ST231 K. pneumoniae strains;
b)comparison of mouse weight between day 1 and day 7 for different ST231 K. pneumoniae strains.
* – p < 0.05, ** – p < 0.01.
The standard strain ATCC® 700603™ as a positive control.
Weight change and survival analysis of mice infected with ST231 Klebsiella pneumoniae. Six-week-old male C57BL/6 mice, grouped in trios, were inoculated intranasally with 30 μl of 108 CFU/ml bacterial suspension. Health monitoring occurred daily for seven days. a)Daily average weight fluctuations among mice infected with different ST231 K. pneumoniae strains; b)comparison of mouse weight between day 1 and day 7 for different ST231 K. pneumoniae strains. * – p < 0.05, ** – p < 0.01. The standard strain ATCC® 700603™ as a positive control.

Fig. 5.

Colony count analysis in mouse alveolar lavage fluids and blood samples post infection.
The growth levels, categorized into low (< 30%), medium (30–60%), and high (> 60%) based on percentage colony formation.
a)The colony counts from cultures of blood samples collected from mice;
b)the colony counts from cultures of alveolar lavage fluids obtained from mice.
Colony count analysis in mouse alveolar lavage fluids and blood samples post infection. The growth levels, categorized into low (< 30%), medium (30–60%), and high (> 60%) based on percentage colony formation. a)The colony counts from cultures of blood samples collected from mice; b)the colony counts from cultures of alveolar lavage fluids obtained from mice.

Fig. 6.

Inflammatory response to ST231 infection in mice.
The levels of key inflammatory markers (CCL2/MCP-1, IL-6, and TNF-α) were quantified in the alveolar lavage fluid (a-c) and blood (d-f) of infected mice. The data from nine independent experiments.
* – p < 0.05, ** – p < 0.01. *** – p < 0.001. The standard strain ATCC® 700603™ as a positive control.
Inflammatory response to ST231 infection in mice. The levels of key inflammatory markers (CCL2/MCP-1, IL-6, and TNF-α) were quantified in the alveolar lavage fluid (a-c) and blood (d-f) of infected mice. The data from nine independent experiments. * – p < 0.05, ** – p < 0.01. *** – p < 0.001. The standard strain ATCC® 700603™ as a positive control.

Fig. 7.

Histopathological impact of ST231 isolates on mouse tissues.
The images, stained with hematoxylin and eosin, reveal significant tissue damage, particularly in the lungs (b–g) and livers (j–o), where extensive cellular infiltration and structural disruption are evident. Renal injury was not significant, and data are not shown.
Histopathological impact of ST231 isolates on mouse tissues. The images, stained with hematoxylin and eosin, reveal significant tissue damage, particularly in the lungs (b–g) and livers (j–o), where extensive cellular infiltration and structural disruption are evident. Renal injury was not significant, and data are not shown.

Clinical and demographic characteristics_

Patient No./isolateAge/sexWardSpecimenDiagnosisInvasive procedureAntimicrobial therapyPrognosis
12(Kp1)52(M)ICUdrainage fluidsabscess of the parotid gland, septic shockincision and drainage of an abscessmeropenem, tigecycline, fluconazoleunknown
32(Kp2)68(M)Cardiac Surgerysputumsevere pneumonia, septic shockheart surgery died
52(Kp3)51(M)Transplantation Centrebloodmalignant tumor of the hilar bile ductresection of hepatic bile duct lesionstigecyclinerecovered
55(Kp4)45(F)Transplantation Centredrainage fluidsliver abscesspartial hepatic lobectomyvancomycin, flucytosinerecovered
206(Kp6)56(M)EICUascitesabdominal infection, infectious shockleft thoracocentesis for drainagecefoperazone sulbactam, polymyxin, meropenem, linezolid, tigecyclineunknown
209(Kp7)78(M)Neurologybloodsepsis, bacteremianon-operativeimipenem, tigecyclinerecovered
255(Kp8)50(M)ICUascitesacute necrotizing pancreatitis, septic shockincision and drainage of retroperitoneal abscessesvancomycin, imipenem, tigecycline, caspofunginrecovered
356(Kp9)68(M)ICUsputumsevere pneumonia, septic shocktracheal tubevancomycin, imipenem, polymyxin, caspofungindied
434(Kp10)37(M)Hepatobiliary Surgerysputumseptic shockabdominal exploratory surgery, tracheal tubeertapenem, meropenem, imipenemunknown
J19(Kp13)58(M)Cardiac Surgerysputumcoronary atherosclerotic heart diseasecoronary bypass graft recovered

Genotypic profiles and resistance mechanisms of Klebsiella pneumoniae isolates_

IsolatesST typePlasmid repliconsSize of plasmid repliconsβ-lactamases and carbapenemases
Kp1ST11IncFIB(K)/IncHI1B, IncR/IncFII, IncFIB, IncFII, ColRNAI, ColKP3193835 bp, 133766 bp, 112792 bp, 84876 bp, 11970 bp, 5596bp, 6141 bpSHV-11, SHV-12, CTX-M-65, LAP-2, OXA-232, KPC-2
Kp2ST231IncFII/IncFIB, IncFIA/IncFII, ColKP3126359 bp, 71307 bp, 6141 bp CTX-M-15, SHV-1, OXA-232CTX-M-186, TEM-1,
Kp3ST231IncFII/IncFIB, IncFIA/IncFII, ColKP3127803 bp, 71307 bp, 6141 bp CTX-M-15, SHV-1, OXA-232CTX-M-186, TEM-1,
Kp4ST231IncFII/IncFIB, IncFIA/IncFII, ColKP3127803 bp, 71688 bp, 6141 bp CTX-M-15, SHV-1, OXA-232CTX-M-186, TEM-1,
Kp6ST231IncFII/IncFIB, IncFIA/IncFII, ColKP3126406 bp, 71307 bp, 6141 bp CTX-M-15, SHV-1, OXA-232CTX-M-186, TEM-1,
Kp7ST231IncFII/IncFIB, IncFIA/IncFII, ColKP3127803 bp, 71307 bp, 6141 bp CTX-M-15, SHV-1, OXA-232CTX-M-186, TEM-1,
Kp8ST231IncFII/IncFIB, IncFIA/IncFII, ColKP3127803 bp, 71307 bp, 6101 bp CTX-M-15, SHV-1, OXA-232CTX-M-186, TEM-1,
Kp9ST231IncFII/IncFIB, IncFIA/IncFII, ColKP3127806 bp, 71307 bp, 6141 bp CTX-M-15, SHV-1, OXA-232CTX-M-186, TEM-1,
Kp10ST231IncFII/IncFIB, IncFIA/IncFII, ColKP3127806bp, 71307 bp, 6141 bpCTX-M-186, TEM-1, CTX-M-15, SHV-1, OXA-232
Kp13ST231IncFIB/IncFII(K), IncFIA/IncFII, ColKP3126359 bp, 71305 bp, 6141 bpCTX-M-186, TEM-1, CTX-M-15, SHV-1, OXA-232
DOI: https://doi.org/10.33073/pjm-2025-007 | Journal eISSN: 2544-4646 | Journal ISSN: 1733-1331
Language: English
Page range: 82 - 94
Submitted on: Nov 12, 2024
Accepted on: Jan 24, 2025
Published on: Mar 26, 2025
Published by: Polish Society of Microbiologists
In partnership with: Paradigm Publishing Services
Publication frequency: 4 issues per year

© 2025 Zhouxun Li, Chunyan Wu, Xuemei Cai, Yongli Song, Xingping Zheng, Yuan He, Guibo Song, published by Polish Society of Microbiologists
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License.