Highly variable effect of sonication to dislodge biofilm-embedded Staphylococcus epidermidis directly quantified by epifluorescence microscopy: an in vitro model study
Abstract Background In cases of prosthetic joint infections, culture of sonication fluid can supplement culture of harvested tissue samples for correct microbial diagnosis. However, discrepant results regarding the increased sensitivity of sonication have been reported in several studies. To what de...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2020-11-01
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| Series: | Journal of Orthopaedic Surgery and Research |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s13018-020-02052-3 |
| _version_ | 1828153041908400128 |
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| author | Erik T. Sandbakken Eivind Witsø Bjørnar Sporsheim Kjartan W. Egeberg Olav A. Foss Linh Hoang Geir Bjerkan Kirsti Løseth Kåre Bergh |
| author_facet | Erik T. Sandbakken Eivind Witsø Bjørnar Sporsheim Kjartan W. Egeberg Olav A. Foss Linh Hoang Geir Bjerkan Kirsti Løseth Kåre Bergh |
| author_sort | Erik T. Sandbakken |
| collection | DOAJ |
| description | Abstract Background In cases of prosthetic joint infections, culture of sonication fluid can supplement culture of harvested tissue samples for correct microbial diagnosis. However, discrepant results regarding the increased sensitivity of sonication have been reported in several studies. To what degree bacteria embedded in biofilm are dislodged during the sonication process has to our knowledge not been fully elucidated. In the present in vitro study, we have evaluated the effect of sonication as a method to dislodge biofilm by quantitative microscopy. Methods We used a standard biofilm method to cover small steel plates with biofilm forming Staphylococcus epidermidis ATCC 35984 and carried out the sonication procedure according to clinical practice. By comparing area covered with biofilm before and after sonication with epifluorescence microscopy, the effect of sonication on biofilm removal was quantified. Two series of experiments were made, one with 24-h biofilm formation and another with 72-h biofilm formation. Confocal laser scanning microscopy (CLSM) and scanning electron microscopy (SEM) were used to confirm whether bacteria were present after sonication. In addition, quantitative bacteriology of sonication fluid was performed. Results Epifluorescence microscopy enabled visualization of biofilm before and after sonication. CLSM and SEM confirmed coccoid cells on the surface after sonication. Biofilm was dislodged in a highly variable manner. Conclusion There is an unexpected high variation seen in the ability of sonication to dislodge biofilm-embedded S. epidermidis in this in vitro model. |
| first_indexed | 2024-04-11T22:19:52Z |
| format | Article |
| id | doaj.art-22828b5e010148e38c70fe5b22b06e57 |
| institution | Directory Open Access Journal |
| issn | 1749-799X |
| language | English |
| last_indexed | 2024-04-11T22:19:52Z |
| publishDate | 2020-11-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Orthopaedic Surgery and Research |
| spelling | doaj.art-22828b5e010148e38c70fe5b22b06e572022-12-22T04:00:12ZengBMCJournal of Orthopaedic Surgery and Research1749-799X2020-11-011511910.1186/s13018-020-02052-3Highly variable effect of sonication to dislodge biofilm-embedded Staphylococcus epidermidis directly quantified by epifluorescence microscopy: an in vitro model studyErik T. Sandbakken0Eivind Witsø1Bjørnar Sporsheim2Kjartan W. Egeberg3Olav A. Foss4Linh Hoang5Geir Bjerkan6Kirsti Løseth7Kåre Bergh8Department of Orthopedic surgery, St Olav’s University HospitalDepartment of Orthopedic surgery, St Olav’s University HospitalCellular and Molecular Imaging Core Facility (CMIC), Norwegian University of Science and TechnologyCellular and Molecular Imaging Core Facility (CMIC), Norwegian University of Science and TechnologyNeuromedicine and Movement Science (INB), Norwegian University of Science and TechnologyCellular and Molecular Imaging Core Facility (CMIC), Norwegian University of Science and TechnologyDepartment of Orthopedic surgery, St Olav’s University HospitalDepartment of Clinical and Molecular Medicine, Norwegian University of Science and TechnologyDepartment of Clinical and Molecular Medicine, Norwegian University of Science and TechnologyAbstract Background In cases of prosthetic joint infections, culture of sonication fluid can supplement culture of harvested tissue samples for correct microbial diagnosis. However, discrepant results regarding the increased sensitivity of sonication have been reported in several studies. To what degree bacteria embedded in biofilm are dislodged during the sonication process has to our knowledge not been fully elucidated. In the present in vitro study, we have evaluated the effect of sonication as a method to dislodge biofilm by quantitative microscopy. Methods We used a standard biofilm method to cover small steel plates with biofilm forming Staphylococcus epidermidis ATCC 35984 and carried out the sonication procedure according to clinical practice. By comparing area covered with biofilm before and after sonication with epifluorescence microscopy, the effect of sonication on biofilm removal was quantified. Two series of experiments were made, one with 24-h biofilm formation and another with 72-h biofilm formation. Confocal laser scanning microscopy (CLSM) and scanning electron microscopy (SEM) were used to confirm whether bacteria were present after sonication. In addition, quantitative bacteriology of sonication fluid was performed. Results Epifluorescence microscopy enabled visualization of biofilm before and after sonication. CLSM and SEM confirmed coccoid cells on the surface after sonication. Biofilm was dislodged in a highly variable manner. Conclusion There is an unexpected high variation seen in the ability of sonication to dislodge biofilm-embedded S. epidermidis in this in vitro model.http://link.springer.com/article/10.1186/s13018-020-02052-3SonicationBiofilm formationStaphylococcus epidermidisFluorescence microscopyConfocal microscopyElectron microscopy |
| spellingShingle | Erik T. Sandbakken Eivind Witsø Bjørnar Sporsheim Kjartan W. Egeberg Olav A. Foss Linh Hoang Geir Bjerkan Kirsti Løseth Kåre Bergh Highly variable effect of sonication to dislodge biofilm-embedded Staphylococcus epidermidis directly quantified by epifluorescence microscopy: an in vitro model study Journal of Orthopaedic Surgery and Research Sonication Biofilm formation Staphylococcus epidermidis Fluorescence microscopy Confocal microscopy Electron microscopy |
| title | Highly variable effect of sonication to dislodge biofilm-embedded Staphylococcus epidermidis directly quantified by epifluorescence microscopy: an in vitro model study |
| title_full | Highly variable effect of sonication to dislodge biofilm-embedded Staphylococcus epidermidis directly quantified by epifluorescence microscopy: an in vitro model study |
| title_fullStr | Highly variable effect of sonication to dislodge biofilm-embedded Staphylococcus epidermidis directly quantified by epifluorescence microscopy: an in vitro model study |
| title_full_unstemmed | Highly variable effect of sonication to dislodge biofilm-embedded Staphylococcus epidermidis directly quantified by epifluorescence microscopy: an in vitro model study |
| title_short | Highly variable effect of sonication to dislodge biofilm-embedded Staphylococcus epidermidis directly quantified by epifluorescence microscopy: an in vitro model study |
| title_sort | highly variable effect of sonication to dislodge biofilm embedded staphylococcus epidermidis directly quantified by epifluorescence microscopy an in vitro model study |
| topic | Sonication Biofilm formation Staphylococcus epidermidis Fluorescence microscopy Confocal microscopy Electron microscopy |
| url | http://link.springer.com/article/10.1186/s13018-020-02052-3 |
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