Optimization of <i>Streptococcus agalactiae</i> Biofilm Culture in a Continuous Flow System for Photoinactivation Studies
<i>Streptococcus agalactiae</i> is a relevant cause of neonatal mortality. It can be transferred to infants via the vaginal tract and cause meningitis, pneumonia, arthritis, or sepsis, among other diseases. The cause of therapy ineffectiveness and infection recurrence is the growth of ba...
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MDPI AG
2021-09-01
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| Series: | Pathogens |
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| Online Access: | https://www.mdpi.com/2076-0817/10/9/1212 |
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| author | Michal K. Pieranski Michal Rychlowski Mariusz Grinholc |
| author_facet | Michal K. Pieranski Michal Rychlowski Mariusz Grinholc |
| author_sort | Michal K. Pieranski |
| collection | DOAJ |
| description | <i>Streptococcus agalactiae</i> is a relevant cause of neonatal mortality. It can be transferred to infants via the vaginal tract and cause meningitis, pneumonia, arthritis, or sepsis, among other diseases. The cause of therapy ineffectiveness and infection recurrence is the growth of bacteria as biofilms. To date, several research teams have attempted to find a suitable medium for the cultivation of <i>S. agalactiae</i> biofilms. Among others, simulated vaginal fluid has been used; however, biofilm production in this medium has been found to be lower than that in tryptic soy broth. We have previously shown that <i>S. agalactiae</i> can be successfully eradicated by photoinactivation in planktonic culture, but there have been no studies on biofilms. The aim of this study was to optimize <i>S. agalactiae</i> biofilm culture conditions to be used in photoinactivation studies. We compared biofilm production by four strains representing the most common serotypes in four different broth media with crystal violet staining. Then, we evaluated stationary biofilm culture in microtiter plates and biofilm growth in a CDC Biofilm Reactor<sup>®</sup> (BioSurface Technologies, Bozeman, MT, USA) under continuous flow conditions. Subsequently, we applied Rose Bengal-mediated photoinactivation to both biofilm models. We have shown that photoinactivation is efficient in biofilm eradication and is not cyto/phototoxic to human keratinocytes. We found conditions allowing for stable and repetitive <i>S. agalactiae</i> biofilm growth in continuous flow conditions, which can be successfully utilized in photoinactivation assays and potentially in all other antibacterial studies. |
| first_indexed | 2024-03-10T07:18:56Z |
| format | Article |
| id | doaj.art-0a0cc3325d23499591bfb099211b7242 |
| institution | Directory Open Access Journal |
| issn | 2076-0817 |
| language | English |
| last_indexed | 2024-03-10T07:18:56Z |
| publishDate | 2021-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Pathogens |
| spelling | doaj.art-0a0cc3325d23499591bfb099211b72422023-11-22T14:43:26ZengMDPI AGPathogens2076-08172021-09-01109121210.3390/pathogens10091212Optimization of <i>Streptococcus agalactiae</i> Biofilm Culture in a Continuous Flow System for Photoinactivation StudiesMichal K. Pieranski0Michal Rychlowski1Mariusz Grinholc2Laboratory of Photobiology and Molecular Diagnostics, Intercollegiate Faculty of Biotechnology University of Gdansk and Medical University of Gdansk, 80-307 Gdansk, PolandLaboratory of Virus Molecular Biology, Intercollegiate Faculty of Biotechnology University of Gdansk and Medical University of Gdansk, 80-307 Gdansk, PolandLaboratory of Photobiology and Molecular Diagnostics, Intercollegiate Faculty of Biotechnology University of Gdansk and Medical University of Gdansk, 80-307 Gdansk, Poland<i>Streptococcus agalactiae</i> is a relevant cause of neonatal mortality. It can be transferred to infants via the vaginal tract and cause meningitis, pneumonia, arthritis, or sepsis, among other diseases. The cause of therapy ineffectiveness and infection recurrence is the growth of bacteria as biofilms. To date, several research teams have attempted to find a suitable medium for the cultivation of <i>S. agalactiae</i> biofilms. Among others, simulated vaginal fluid has been used; however, biofilm production in this medium has been found to be lower than that in tryptic soy broth. We have previously shown that <i>S. agalactiae</i> can be successfully eradicated by photoinactivation in planktonic culture, but there have been no studies on biofilms. The aim of this study was to optimize <i>S. agalactiae</i> biofilm culture conditions to be used in photoinactivation studies. We compared biofilm production by four strains representing the most common serotypes in four different broth media with crystal violet staining. Then, we evaluated stationary biofilm culture in microtiter plates and biofilm growth in a CDC Biofilm Reactor<sup>®</sup> (BioSurface Technologies, Bozeman, MT, USA) under continuous flow conditions. Subsequently, we applied Rose Bengal-mediated photoinactivation to both biofilm models. We have shown that photoinactivation is efficient in biofilm eradication and is not cyto/phototoxic to human keratinocytes. We found conditions allowing for stable and repetitive <i>S. agalactiae</i> biofilm growth in continuous flow conditions, which can be successfully utilized in photoinactivation assays and potentially in all other antibacterial studies.https://www.mdpi.com/2076-0817/10/9/1212<i>S. agalactiae</i>Group B <i>Streptococcus</i> (GBS)biofilmcontinuous flowCenter for Disease Control and Prevention (CDC) Biofilm Reactorphotoinactivation |
| spellingShingle | Michal K. Pieranski Michal Rychlowski Mariusz Grinholc Optimization of <i>Streptococcus agalactiae</i> Biofilm Culture in a Continuous Flow System for Photoinactivation Studies Pathogens <i>S. agalactiae</i> Group B <i>Streptococcus</i> (GBS) biofilm continuous flow Center for Disease Control and Prevention (CDC) Biofilm Reactor photoinactivation |
| title | Optimization of <i>Streptococcus agalactiae</i> Biofilm Culture in a Continuous Flow System for Photoinactivation Studies |
| title_full | Optimization of <i>Streptococcus agalactiae</i> Biofilm Culture in a Continuous Flow System for Photoinactivation Studies |
| title_fullStr | Optimization of <i>Streptococcus agalactiae</i> Biofilm Culture in a Continuous Flow System for Photoinactivation Studies |
| title_full_unstemmed | Optimization of <i>Streptococcus agalactiae</i> Biofilm Culture in a Continuous Flow System for Photoinactivation Studies |
| title_short | Optimization of <i>Streptococcus agalactiae</i> Biofilm Culture in a Continuous Flow System for Photoinactivation Studies |
| title_sort | optimization of i streptococcus agalactiae i biofilm culture in a continuous flow system for photoinactivation studies |
| topic | <i>S. agalactiae</i> Group B <i>Streptococcus</i> (GBS) biofilm continuous flow Center for Disease Control and Prevention (CDC) Biofilm Reactor photoinactivation |
| url | https://www.mdpi.com/2076-0817/10/9/1212 |
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