68b88321a66fd941a77e7b7c97c45c36 300797.pdf 914c990765f5c97546db4262d1f7eca9d852cd47 300797.pdf 1c7c42fc36ed0b68954f1d3f0cbff9ab0902c2343a29ce6628e833c9fcb2bd6b 300797.pdf Title: In Vitro Effect of Three-Antibiotic Combinations plus Potential Antibiofilm Agents against Biofilm-Producing Mycobacterium avium and Mycobacterium intracellulare Clinical Isolates Subject: Patients with chronic pulmonary diseases infected by Mycobacterium avium complex (MAC) often develop complications and suffer from treatment failure due to biofilm formation. There is a lack of correlation between in vitro susceptibility tests and the treatment of clinical isolates producing biofilm. We performed susceptibility tests of 10 different three-drug combinations, including two recommended in the guidelines, in biofilm forms of eight MAC clinical isolates. Biofilm developed in the eight isolates following incubation of the inoculum for 3 weeks. Then, the biofilm was treated with three-drug combinations with and without the addition of potential antibiofilm agents (PAAs). Biofilm bactericidal concentrations (BBCs) were determined using the Vizion lector system. All selected drug combinations showed synergistic activity, reducing BBC values compared to those treated with single drugs, but BBC values remained high enough to treat patients. However, with the addition of PAAs, the BBCs steadily decreased, achieving similar values to the combinations in planktonic forms and showing synergistic activity in all the combinations and in both species. In conclusion, three-drug combinations with PAAs showed synergistic activity in biofilm forms of MAC isolates. Our results suggest the need for clinical studies introducing PAAs combined with antibiotics for the treatment of patients with pulmonary diseases infected by MAC. Keywords: biofilm; antibiotic combinations; Mycobacterium avium complex; antibiotics; antibiofilm agents Author: Sara Batista, Mariana Fernandez-Pittol, Lorena San Nicolás, Diego Martínez, Marc Rubio, Montserrat Garrigo, Jordi Vila, Griselda Tudó and Julian González-Martin Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Wed Sep 6 06:05:18 2023 CEST ModDate: Wed Sep 6 06:07:21 2023 CEST Custom Metadata: no Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 15 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 2505174 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- YVCPHT+URWPalladioL-Bold Type 1 Custom yes yes no 10 0 QQXUHG+URWPalladioL-Roma Type 1 Custom yes yes no 15 0 UGALXF+URWPalladioL-Ital Type 1 Custom yes yes no 19 0 JVWPGJ+URWPalladioL-BoldItal Type 1 Custom yes yes no 23 0 HEEXOW+VnURWPalladioL Type 1 Custom yes yes no 27 0 PRJKGQ+VnURWPalladioL-Bold Type 1 Custom yes yes no 32 0 YYFPCJ+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 51 0 DHYYSE+CMSY10 Type 1 Builtin yes yes no 76 0 CAMXTK+EURM10 Type 1 Builtin yes yes no 80 0 XIRVNA+EURB10 Type 1 Builtin yes yes no 90 0 NOEACM+PalatinoLinotype TrueType MacRoman yes yes no 116 0 NOEDGM+PalatinoLinotype,Italic TrueType MacRoman yes yes no 119 0 NODPFJ+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 122 0 NODPFK+PalatinoLinotype CID TrueType Identity-H yes yes yes 125 0 NODPFL+PalatinoLinotype TrueType WinAnsi yes yes no 131 0 NODPFN+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 134 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2024-09-20 02:51:01 CEST RepresentationInformation: 300797.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2024-09-19 10:07:07 CEST Size: 2505174 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 356 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: section.1 Item: Title: Results Destination: section.2 Item: Title: Discussion Destination: section.3 Item: Title: Materials and Methods Destination: section.4 Children: Item: Title: Mycobacterium avium Intracellulare Complex Isolates Destination: subsection.4.1 Item: Title: Antibiotics and Potential Antibiofilm Agents (PAAs) Destination: subsection.4.2 Item: Title: Antimicrobial Susceptibility Testing in Planktonic Forms Destination: subsection.4.3 Item: Title: Biofilm Formation in Clinical Isolates Destination: subsection.4.4 Item: Title: Biofilm Treatment Study Destination: subsection.4.5 Children: Item: Title: Antibiotic Alone plus PAAs in Biofilm Destination: subsubsection.4.5.1 Item: Title: Determination of Biofilm Bactericidal Concentration Destination: subsubsection.4.5.2 Item: Title: Determination of the Fractional Inhibitory Concentration Index Destination: subsubsection.4.5.3 Item: Title: Conclusions Destination: section.5 Item: Title: References Destination: section.A. Info: Title: In Vitro Effect of Three-Antibiotic Combinations plus Potential Antibiofilm Agents against Biofilm-Producing Mycobacterium avium and Mycobacterium intracellulare Clinical Isolates Author: Sara Batista, Mariana Fernandez-Pittol, Lorena San Nicolás, Diego Martínez, Marc Rubio, Montserrat Garrigo, Jordi Vila, Griselda Tudó and Julian González-Martin Subject: Patients with chronic pulmonary diseases infected by Mycobacterium avium complex (MAC) often develop complications and suffer from treatment failure due to biofilm formation. There is a lack of correlation between in vitro susceptibility tests and the treatment of clinical isolates producing biofilm. We performed susceptibility tests of 10 different three-drug combinations, including two recommended in the guidelines, in biofilm forms of eight MAC clinical isolates. Biofilm developed in the eight isolates following incubation of the inoculum for 3 weeks. Then, the biofilm was treated with three-drug combinations with and without the addition of potential antibiofilm agents (PAAs). Biofilm bactericidal concentrations (BBCs) were determined using the Vizion lector system. All selected drug combinations showed synergistic activity, reducing BBC values compared to those treated with single drugs, but BBC values remained high enough to treat patients. However, with the addition of PAAs, the BBCs steadily decreased, achieving similar values to the combinations in planktonic forms and showing synergistic activity in all the combinations and in both species. In conclusion, three-drug combinations with PAAs showed synergistic activity in biofilm forms of MAC isolates. Our results suggest the need for clinical studies introducing PAAs combined with antibiotics for the treatment of patients with pulmonary diseases infected by MAC. 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