a882a35acbd1a000296d91c0e27808cc antibiotics-14-00724.pdf 3551f191403fec142c31c6f881452893e2f184e4 antibiotics-14-00724.pdf 7633684ffc81dba2dbf9ca4e33d130be57595d85cbcb63af17788815ad0c361c antibiotics-14-00724.pdf Title: Exploring the Microbiome of Diabetic Foot Ulcers: A Focus on Cases with a Clinical Worse Outcome Subject: Background/Objectives: We evaluated the diabetic foot ulcer (DFU) microbiome in clinical situations identified as risk factors for a worse outcome and explored the roles of the most abundant microorganisms. Methods: A prospective multicenter cohort of diabetic patients with DFU were followed up for 6 months. We obtained a DFU tissue biopsy for microbiome analysis at the baseline visit. Genomic DNA was extracted (QIAamp DNA Mini Kit, Qiagen, Hilden, Germany) and quantified (QuantiFluor dsDNA System, Promega, Madison, WI, USA), with analysis of bacterial communities focusing on relative abundances (RA) and on alpha and beta diversity. Results: Overall, 59 DFUs were analyzed. DFUs of long duration (4 weeks) presented a higher RA of Gammaproteobacteria compared with ulcers of short duration (p = 0.02). Non-infected DFUs had a higher proportion of Actinobacteriota phyla than infected DFUs and, particularly, a higher RA of Corynebacterium genera (means SD: 0.063 0.14 vs. 0.028 0.13, respectively; p = 0.03). Regarding the pathogenic role of Staphylococcus aureus, DFUs with low S. aureus bacterial loads (<106 CFU/mL) compared with those with high loads (106 CFU/mL) showed a higher Corynebacterium RA (0.045 0.08 vs. 0.003 0.01, respectively; p = 0.01). Conclusions: In clinical situations associated with poor DFU outcomes, we observed a predominance of Gammaproteobacteria in the microbiome of long-duration ulcers and a higher RA of Corynebacterium in non-infected DFUs. An inverse relationship between the predominance of Corynebacterium and the S. aureus bacterial load in DFUs was also noted, which may suggest these commensals have a modulatory role. Further studies should explore the clinical utility of microbiome analysis for DFUs. Keywords: microbiome; chronic diabetic foot ulcers; Gammaproteobacteria Author: Laura Soldevila-Boixader, Anna Carrera-Salinas, Isabel Mur, Laura Morata, Alba Rivera, Jordi Bosch, Abelardo Montero-Saez, Jéssica Martínez Castillejo, Natividad Benito, Sara Martí and Oscar Murillo Creator: LaTeX with hyperref Producer: pdfTeX-1.40.25; modified using OpenPDF 1.4.2 CreationDate: Fri Jul 18 16:08:49 2025 CEST ModDate: Fri Jul 18 16:11:56 2025 CEST Custom Metadata: yes Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 14 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 2020737 bytes Optimized: no PDF version: 1.5 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- CRKHEV+VnURWPalladioL Type 1 Custom yes yes yes 77 0 FCEUIH+URWPalladioL-Roma Type 1 Custom yes yes yes 78 0 IYMDAR+URWPalladioL-Ital Type 1 Custom yes yes yes 79 0 EVGJSF+URWPalladioL-Bold Type 1 Custom yes yes yes 80 0 BCGYZM+CMSY10 Type 1 Builtin yes yes yes 81 0 WDTPMW+URWPalladioL-BoldItal Type 1 Custom yes yes yes 140 0 LEFFJM+PalatinoLinotype-Italic CID TrueType Identity-H yes yes yes 151 0 LEFFGL+PalatinoLinotype-Bold CID TrueType Identity-H yes yes yes 152 0 LEFFIL+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 153 0 LEFFJM+PalatinoLinotype-Italic CID TrueType Identity-H yes yes yes 162 0 LEFFGL+PalatinoLinotype-Bold CID TrueType Identity-H yes yes yes 163 0 LEFFIL+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 164 0 RIJDHR+CMBSY10 Type 1 Builtin yes yes yes 232 0 LEFFJM+PalatinoLinotype-Italic CID TrueType Identity-H yes yes yes 198 0 LEFFGL+PalatinoLinotype-Bold CID TrueType Identity-H yes yes yes 199 0 LEFFIL+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 200 0 LEFFJM+PalatinoLinotype-Italic CID TrueType Identity-H yes yes yes 225 0 LEFFGL+PalatinoLinotype-Bold CID TrueType Identity-H yes yes yes 226 0 LEFFIL+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 227 0 LEFJNH+PalatinoLinotype-BoldItalic CID TrueType Identity-H yes yes yes 228 0 OVNEXW+EURM10 Type 1 Builtin yes yes yes 343 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2025-10-18 03:54:15 CEST RepresentationInformation: antibiotics-14-00724.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2025-10-17 19:25:49 CEST Size: 2020737 Format: PDF Version: 1.5 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 435 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: ViewerPreferences: HideToolbar: false HideMenubar: false HideWindowUI: false FitWindow: true CenterWindow: false DisplayDocTitle: false NonFullScreenPageMode: UseNone Direction: L2R ViewArea: CropBox ViewClip: CropBox PrintArea: CropBox PageClip: CropBox PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Item: Title: Results Item: Title: Discussion Item: Title: Materials and Methods Children: Item: Title: Study Design, Setting, and Patients Item: Title: Definitions, Data Collection, and Clinical Management Item: Title: Microbiological Studies and Statistical Analyses Item: Title: Conclusions Item: Title: References Info: Title: Exploring the Microbiome of Diabetic Foot Ulcers: A Focus on Cases with a Clinical Worse Outcome Author: Laura Soldevila-Boixader, Anna Carrera-Salinas, Isabel Mur, Laura Morata, Alba Rivera, Jordi Bosch, Abelardo Montero-Saez, Jéssica Martínez Castillejo, Natividad Benito, Sara Martí and Oscar Murillo Subject: Background/Objectives: We evaluated the diabetic foot ulcer (DFU) microbiome in clinical situations identified as risk factors for a worse outcome and explored the roles of the most abundant microorganisms. Methods: A prospective multicenter cohort of diabetic patients with DFU were followed up for 6 months. We obtained a DFU tissue biopsy for microbiome analysis at the baseline visit. Genomic DNA was extracted (QIAamp DNA Mini Kit, Qiagen, Hilden, Germany) and quantified (QuantiFluor dsDNA System, Promega, Madison, WI, USA), with analysis of bacterial communities focusing on relative abundances (RA) and on alpha and beta diversity. Results: Overall, 59 DFUs were analyzed. DFUs of long duration (4 weeks) presented a higher RA of Gammaproteobacteria compared with ulcers of short duration (p = 0.02). Non-infected DFUs had a higher proportion of Actinobacteriota phyla than infected DFUs and, particularly, a higher RA of Corynebacterium genera (means SD: 0.063 0.14 vs. 0.028 0.13, respectively; p = 0.03). Regarding the pathogenic role of Staphylococcus aureus, DFUs with low S. aureus bacterial loads (<106 CFU/mL) compared with those with high loads (106 CFU/mL) showed a higher Corynebacterium RA (0.045 0.08 vs. 0.003 0.01, respectively; p = 0.01). Conclusions: In clinical situations associated with poor DFU outcomes, we observed a predominance of Gammaproteobacteria in the microbiome of long-duration ulcers and a higher RA of Corynebacterium in non-infected DFUs. An inverse relationship between the predominance of Corynebacterium and the S. aureus bacterial load in DFUs was also noted, which may suggest these commensals have a modulatory role. Further studies should explore the clinical utility of microbiome analysis for DFUs. Keywords: microbiome; chronic diabetic foot ulcers; Gammaproteobacteria Creator: LaTeX with hyperref Producer: pdfTeX-1.40.25; modified using OpenPDF 1.4.2 CreationDate: Fri Jul 18 16:08:49 CEST 2025 ModDate: Fri Jul 18 16:11:56 CEST 2025 ID: 0xb6b0904545a75ac684c10c06f2376e7c, 0x1fa8703c9b813d44433adfc3fc436b56 Filters: FilterPipeline: FlateDecode Fonts: Type1: Font: BaseFont: EVGJSF+URWPalladioL-Bold FontSubset: true FirstChar: 3 LastChar: 237 FontDescriptor: FontName: EVGJSF+URWPalladioL-Bold Flags: Symbolic FontBBox: -152, -301, 1000, 935 FontFile: true EncodingDictionary: Differences: true ToUnicode: true Font: BaseFont: BCGYZM+CMSY10 FontSubset: true FirstChar: 2 LastChar: 48 FontDescriptor: FontName: BCGYZM+CMSY10 Flags: Symbolic FontBBox: -29, -960, 1116, 775 FontFile: true ToUnicode: true Font: BaseFont: OVNEXW+EURM10 FontSubset: true FirstChar: 22 LastChar: 22 FontDescriptor: FontName: OVNEXW+EURM10 Flags: Symbolic FontBBox: -32, -243, 1060, 720 FontFile: true ToUnicode: true Font: BaseFont: RIJDHR+CMBSY10 FontSubset: true FirstChar: 21 LastChar: 21 FontDescriptor: FontName: RIJDHR+CMBSY10 Flags: Symbolic FontBBox: -26, -940, 1332, 825 FontFile: true ToUnicode: true Font: BaseFont: WDTPMW+URWPalladioL-BoldItal FontSubset: true FirstChar: 46 LastChar: 117 FontDescriptor: FontName: WDTPMW+URWPalladioL-BoldItal Flags: Symbolic FontBBox: -170, -300, 1073, 935 FontFile: true EncodingDictionary: Differences: true ToUnicode: true Font: BaseFont: CRKHEV+VnURWPalladioL FontSubset: true FirstChar: 146 LastChar: 177 FontDescriptor: FontName: CRKHEV+VnURWPalladioL Flags: Symbolic FontBBox: -166, -283, 1021, 943 FontFile: true EncodingDictionary: Differences: true ToUnicode: true Font: BaseFont: FCEUIH+URWPalladioL-Roma FontSubset: true FirstChar: 1 LastChar: 252 FontDescriptor: FontName: FCEUIH+URWPalladioL-Roma Flags: Symbolic FontBBox: -166, -283, 1021, 943 FontFile: true EncodingDictionary: Differences: true ToUnicode: true Font: BaseFont: IYMDAR+URWPalladioL-Ital FontSubset: true FirstChar: 2 LastChar: 150 FontDescriptor: FontName: IYMDAR+URWPalladioL-Ital Flags: Symbolic FontBBox: -170, -305, 1010, 941 FontFile: true EncodingDictionary: Differences: true ToUnicode: true Pages: Page: Sequence: 1 Annotations: Annotation: Subtype: Link Rect: 110, 177, 152, 186 Annotation: Subtype: Link Rect: 34, 165, 147, 174 Annotation: Subtype: Link Rect: 36, 75, 152, 84 Annotation: Subtype: Link Rect: 34, 63, 89, 72 Annotation: Subtype: Link Rect: 34, 773, 151, 809 Annotation: Subtype: Link Rect: 515, 773, 560, 809 Annotation: Subtype: Link Rect: 170, 664, 181, 677 Annotation: Subtype: Link Rect: 436, 664, 447, 677 Annotation: Subtype: Link Rect: 211, 650, 222, 663 Annotation: Subtype: Link Rect: 354, 650, 365, 663 Annotation: Subtype: Link Rect: 479, 650, 491, 663 Annotation: Subtype: Link Rect: 399, 27, 560, 37 Annotation: Subtype: Link Rect: 34, 344, 87, 363 Page: Sequence: 2 Annotations: Annotation: Subtype: Link Rect: 400, 552, 407, 560 ActionDest: 41 Annotation: Subtype: Link Rect: 408, 552, 415, 560 ActionDest: 41 Annotation: Subtype: Link Rect: 456, 523, 463, 533 ActionDest: 41 Annotation: Subtype: Link Rect: 463, 524, 470, 533 ActionDest: 128 Annotation: Subtype: Link Rect: 319, 426, 326, 435 ActionDest: 41 Annotation: Subtype: Link Rect: 326, 426, 333, 435 ActionDest: 128 Annotation: Subtype: Link Rect: 334, 426, 341, 435 ActionDest: 128 Annotation: Subtype: Link Rect: 196, 370, 203, 379 ActionDest: 128 Annotation: Subtype: Link Rect: 203, 370, 210, 379 ActionDest: 128 Annotation: Subtype: Link Rect: 384, 287, 391, 296 ActionDest: 128 Annotation: Subtype: Link Rect: 190, 172, 197, 184 ActionDest: 101 Page: Sequence: 3 Annotations: Annotation: Subtype: Link Rect: 541, 479, 548, 491 ActionDest: 102 Annotation: Subtype: Link Rect: 428, 451, 435, 463 ActionDest: 102 Annotation: Subtype: Link Rect: 386, 395, 393, 407 ActionDest: 103 Annotation: Subtype: Link Rect: 407, 144, 414, 156 ActionDest: 104 Page: Sequence: 4 Page: Sequence: 5 Page: Sequence: 6 Annotations: Annotation: Subtype: Link Rect: 360, 81, 367, 93 ActionDest: 254 Page: Sequence: 7 Page: Sequence: 8 Annotations: Annotation: Subtype: Link Rect: 196, 616, 208, 625 ActionDest: 128 Annotation: Subtype: Link Rect: 208, 616, 220, 625 ActionDest: 128 Annotation: Subtype: Link Rect: 297, 588, 309, 597 ActionDest: 128 Annotation: Subtype: Link Rect: 310, 588, 322, 597 ActionDest: 128 Annotation: Subtype: Link Rect: 487, 435, 499, 444 ActionDest: 128 Annotation: Subtype: Link Rect: 251, 421, 263, 430 ActionDest: 128 Annotation: Subtype: Link Rect: 412, 393, 419, 402 ActionDest: 128 Annotation: Subtype: Link Rect: 419, 393, 431, 402 ActionDest: 128 Annotation: Subtype: Link Rect: 432, 393, 444, 402 ActionDest: 128 Annotation: Subtype: Link Rect: 469, 338, 481, 347 ActionDest: 128 Annotation: Subtype: Link Rect: 468, 212, 480, 221 ActionDest: 128 Page: Sequence: 9 Annotations: Annotation: Subtype: Link Rect: 529, 605, 541, 614 ActionDest: 128 Annotation: Subtype: Link Rect: 544, 605, 556, 614 ActionDest: 128 Annotation: Subtype: Link Rect: 425, 494, 437, 503 ActionDest: 128 Annotation: Subtype: Link Rect: 228, 424, 240, 433 ActionDest: 128 Annotation: Subtype: Link Rect: 235, 410, 247, 419 ActionDest: 128 Annotation: Subtype: Link Rect: 247, 410, 259, 419 ActionDest: 128 Annotation: Subtype: Link Rect: 544, 327, 556, 336 ActionDest: 128 Page: Sequence: 10 Annotations: Annotation: Subtype: Link Rect: 258, 450, 265, 459 ActionDest: 128 Annotation: Subtype: Link Rect: 258, 394, 270, 403 ActionDest: 128 Annotation: Subtype: Link Rect: 270, 394, 282, 403 ActionDest: 128 Annotation: Subtype: Link Rect: 387, 366, 399, 375 ActionDest: 333 Annotation: Subtype: Link Rect: 419, 311, 426, 320 ActionDest: 128 Annotation: Subtype: Link Rect: 443, 269, 455, 278 ActionDest: 333 Annotation: Subtype: Link Rect: 456, 269, 468, 278 ActionDest: 333 Annotation: Subtype: Link Rect: 374, 187, 381, 196 ActionDest: 128 Page: Sequence: 11 Annotations: Annotation: Subtype: Link Rect: 261, 633, 273, 642 ActionDest: 333 Annotation: Subtype: Link Rect: 412, 577, 424, 586 ActionDest: 333 Annotation: Subtype: Link Rect: 544, 508, 556, 517 ActionDest: 333 Annotation: Subtype: Link Rect: 451, 438, 463, 447 ActionDest: 333 Annotation: Subtype: Link Rect: 476, 173, 488, 182 ActionDest: 333 Annotation: Subtype: Link Rect: 337, 115, 560, 127 Annotation: Subtype: Link Rect: 165, 101, 228, 113 Page: Sequence: 12 Annotations: Annotation: Subtype: Link Rect: 526, 533, 560, 544 Annotation: Subtype: Link Rect: 165, 519, 390, 530 Annotation: Subtype: Link Rect: 499, 136, 537, 147 Annotation: Subtype: Link Rect: 301, 111, 339, 122 Annotation: Subtype: Link Rect: 389, 85, 427, 96 Page: Sequence: 13 Annotations: Annotation: Subtype: Link Rect: 427, 743, 464, 754 Annotation: Subtype: Link Rect: 210, 706, 247, 716 Annotation: Subtype: Link Rect: 122, 680, 159, 690 Annotation: Subtype: Link Rect: 131, 655, 168, 665 Annotation: Subtype: Link Rect: 174, 655, 210, 665 Annotation: Subtype: Link Rect: 519, 628, 557, 639 Annotation: Subtype: Link Rect: 59, 616, 95, 627 Annotation: Subtype: Link Rect: 227, 578, 265, 588 Annotation: Subtype: Link Rect: 271, 578, 307, 588 Annotation: Subtype: Link Rect: 59, 553, 96, 563 Annotation: Subtype: Link Rect: 519, 539, 557, 550 Annotation: Subtype: Link Rect: 170, 501, 207, 512 Annotation: Subtype: Link Rect: 213, 501, 249, 512 Annotation: Subtype: Link Rect: 362, 475, 400, 486 Annotation: Subtype: Link Rect: 406, 475, 442, 486 Annotation: Subtype: Link Rect: 504, 450, 541, 461 Annotation: Subtype: Link Rect: 448, 425, 485, 435 Annotation: Subtype: Link Rect: 321, 399, 358, 410 Annotation: Subtype: Link Rect: 99, 374, 137, 384 Annotation: Subtype: Link Rect: 520, 347, 557, 358 Annotation: Subtype: Link Rect: 264, 322, 301, 333 Annotation: Subtype: Link Rect: 151, 285, 189, 295 Annotation: Subtype: Link Rect: 253, 258, 291, 269 Annotation: Subtype: Link Rect: 471, 233, 509, 244 Annotation: Subtype: Link Rect: 515, 233, 551, 244 Annotation: Subtype: Link Rect: 240, 195, 278, 205 Annotation: Subtype: Link Rect: 284, 195, 320, 205 Annotation: Subtype: Link Rect: 375, 169, 412, 180 Annotation: Subtype: Link Rect: 418, 169, 455, 180 Annotation: Subtype: Link Rect: 327, 143, 365, 154 Annotation: Subtype: Link Rect: 297, 106, 334, 116 Annotation: Subtype: Link Rect: 59, 80, 96, 90 Page: Sequence: 14 Annotations: Annotation: Subtype: Link Rect: 258, 705, 295, 716 Annotation: Subtype: Link Rect: 265, 667, 303, 678 Annotation: Subtype: Link Rect: 307, 641, 344, 652 Annotation: Subtype: Link Rect: 486, 616, 523, 627 Annotation: Subtype: Link Rect: 189, 591, 227, 601 Annotation: Subtype: Link Rect: 236, 565, 274, 576 Annotation: Subtype: Link Rect: 280, 565, 316, 576 Checksum: 1940ad04 Type: CRC32 Checksum: a882a35acbd1a000296d91c0e27808cc Type: MD5 Checksum: 3551f191403fec142c31c6f881452893e2f184e4 Type: SHA-1 Checksum: 7633684ffc81dba2dbf9ca4e33d130be57595d85cbcb63af17788815ad0c361c Type: SHA-256