A new spray‐based method for the in‐vitro development of dry‐surface biofilms
Abstract The inanimate environment immediately surrounding the patient in healthcare facilities is a reservoir of microorganisms embedded in dry‐surface biofilms (DSB). These biofilms, first highlighted in 2012, are increasingly studied, but currently available in‐vitro models only allow for the gro...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Wiley
2023-02-01
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| Series: | MicrobiologyOpen |
| Subjects: | |
| Online Access: | https://doi.org/10.1002/mbo3.1330 |
| _version_ | 1797895064556404736 |
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| author | Esther Christine Claude Olive Myriam Louisin Moustapha Dramé Karine Marion‐Sanchez |
| author_facet | Esther Christine Claude Olive Myriam Louisin Moustapha Dramé Karine Marion‐Sanchez |
| author_sort | Esther Christine |
| collection | DOAJ |
| description | Abstract The inanimate environment immediately surrounding the patient in healthcare facilities is a reservoir of microorganisms embedded in dry‐surface biofilms (DSB). These biofilms, first highlighted in 2012, are increasingly studied, but currently available in‐vitro models only allow for the growth of semi‐hydrated biofilms. We developed a new in‐vitro method under actual dehydration conditions based on the hypothesis that surface contamination is mainly due to splashes of respiratory secretions. The main objective of this study was to show that the operating conditions we have defined allowed the growth of DSB with a methicillin resistant Staphylococcus aureus strain. The second objective was to show that extended‐spectrum beta‐lactamase‐producing Enterobacteriaceae, that is, Klebsiella pneumoniae and Enterobacter cloacae were also able to grow such biofilms under these conditions. Monobacterial suspensions in sterile artificial saliva (SAS) were sprayed onto polyethylene surfaces. Nutrients and hydration were provided daily by spraying SAS enriched with 20% of Brain Heart Infusion broth. The primary outcome was mean surface coverage measured by image analysis after crystal violet staining. The method applied to S. aureus for 12 days resulted in reproducible and repeatable DSB consisting of isolated and confluent microcolonies embedded in extracellular polymeric substances as shown in scanning electron microscopy images. Similar DSB were obtained with both Enterobacteriaceae applying the same method. No interspecies variation was shown between the three strains in terms of surface coverage. These first trials are the starting point for a 3‐year study currently in process. |
| first_indexed | 2024-04-10T07:19:23Z |
| format | Article |
| id | doaj.art-2c1eb5dcc7784771befea1f8b2e89134 |
| institution | Directory Open Access Journal |
| issn | 2045-8827 |
| language | English |
| last_indexed | 2024-04-10T07:19:23Z |
| publishDate | 2023-02-01 |
| publisher | Wiley |
| record_format | Article |
| series | MicrobiologyOpen |
| spelling | doaj.art-2c1eb5dcc7784771befea1f8b2e891342023-02-24T13:44:33ZengWileyMicrobiologyOpen2045-88272023-02-01121n/an/a10.1002/mbo3.1330A new spray‐based method for the in‐vitro development of dry‐surface biofilmsEsther Christine0Claude Olive1Myriam Louisin2Moustapha Dramé3Karine Marion‐Sanchez4Department of Bacteriology Hygiene and Environment Laboratory, CHU Martinique CS 90632 Fort‐de‐France Cedex MartiniqueDepartment of Bacteriology Hygiene and Environment Laboratory, CHU Martinique CS 90632 Fort‐de‐France Cedex MartiniqueDepartment of Bacteriology Hygiene and Environment Laboratory, CHU Martinique CS 90632 Fort‐de‐France Cedex MartiniqueDepartment of Clinical Research and Innovation CHU Martinique CS 90632 Fort‐de‐France Cedex MartiniqueDepartment of Bacteriology Hygiene and Environment Laboratory, CHU Martinique CS 90632 Fort‐de‐France Cedex MartiniqueAbstract The inanimate environment immediately surrounding the patient in healthcare facilities is a reservoir of microorganisms embedded in dry‐surface biofilms (DSB). These biofilms, first highlighted in 2012, are increasingly studied, but currently available in‐vitro models only allow for the growth of semi‐hydrated biofilms. We developed a new in‐vitro method under actual dehydration conditions based on the hypothesis that surface contamination is mainly due to splashes of respiratory secretions. The main objective of this study was to show that the operating conditions we have defined allowed the growth of DSB with a methicillin resistant Staphylococcus aureus strain. The second objective was to show that extended‐spectrum beta‐lactamase‐producing Enterobacteriaceae, that is, Klebsiella pneumoniae and Enterobacter cloacae were also able to grow such biofilms under these conditions. Monobacterial suspensions in sterile artificial saliva (SAS) were sprayed onto polyethylene surfaces. Nutrients and hydration were provided daily by spraying SAS enriched with 20% of Brain Heart Infusion broth. The primary outcome was mean surface coverage measured by image analysis after crystal violet staining. The method applied to S. aureus for 12 days resulted in reproducible and repeatable DSB consisting of isolated and confluent microcolonies embedded in extracellular polymeric substances as shown in scanning electron microscopy images. Similar DSB were obtained with both Enterobacteriaceae applying the same method. No interspecies variation was shown between the three strains in terms of surface coverage. These first trials are the starting point for a 3‐year study currently in process.https://doi.org/10.1002/mbo3.1330artificial salivadry‐surface biofilmEnterobacter cloacaeKlebsiella pneumoniaeStaphylococcus aureus |
| spellingShingle | Esther Christine Claude Olive Myriam Louisin Moustapha Dramé Karine Marion‐Sanchez A new spray‐based method for the in‐vitro development of dry‐surface biofilms MicrobiologyOpen artificial saliva dry‐surface biofilm Enterobacter cloacae Klebsiella pneumoniae Staphylococcus aureus |
| title | A new spray‐based method for the in‐vitro development of dry‐surface biofilms |
| title_full | A new spray‐based method for the in‐vitro development of dry‐surface biofilms |
| title_fullStr | A new spray‐based method for the in‐vitro development of dry‐surface biofilms |
| title_full_unstemmed | A new spray‐based method for the in‐vitro development of dry‐surface biofilms |
| title_short | A new spray‐based method for the in‐vitro development of dry‐surface biofilms |
| title_sort | new spray based method for the in vitro development of dry surface biofilms |
| topic | artificial saliva dry‐surface biofilm Enterobacter cloacae Klebsiella pneumoniae Staphylococcus aureus |
| url | https://doi.org/10.1002/mbo3.1330 |
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