MBRA-2: a Modified Chemostat System to Culture Biofilms
ABSTRACT Culture-dependent approaches for investigating microbial ecology aim to model the nutrient content of specific environments by simplifying the system for high-resolution molecular analysis. These in vitro systems are enticing due to their increased throughput compared to animal models, flex...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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American Society for Microbiology
2023-02-01
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| Series: | Microbiology Spectrum |
| Subjects: | |
| Online Access: | https://journals.asm.org/doi/10.1128/spectrum.02928-22 |
| _version_ | 1811164402639962112 |
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| author | Justin N. Jens Daniel J. Breiner Rachel L. Neve Matilda M. Fiebig Vanessa V. Phelan |
| author_facet | Justin N. Jens Daniel J. Breiner Rachel L. Neve Matilda M. Fiebig Vanessa V. Phelan |
| author_sort | Justin N. Jens |
| collection | DOAJ |
| description | ABSTRACT Culture-dependent approaches for investigating microbial ecology aim to model the nutrient content of specific environments by simplifying the system for high-resolution molecular analysis. These in vitro systems are enticing due to their increased throughput compared to animal models, flexibility in modulating nutrient content and community composition, scaling of culture volume to isolate biological molecules, and control of environmental parameters, such as temperature, humidity, and nutrient flow. However, different devices are used to investigate homogenous, planktonic microbial communities and heterogeneous biofilms. Here, we present the minibioreactor array 2 (MBRA-2) with media rails, a benchtop multireactor system derived from the MBRA system that enables researchers to use the same system to grow planktonic and biofilm cultures. We simplified flow through the system and reduced contamination, leakage, and time required for array assembly by designing and implementing a reusable media rail to replace the branched tubing traditionally used to convey media through chemostat arrays. Additionally, we altered the structure of the six-bioreactor strip to incorporate a removable lid to provide easy access to the bioreactor wells, enabling biofilm recovery and thorough cleaning for reuse. Using Pseudomonas aeruginosa, a model biofilm-producing organism, we show that the technical improvements of the MBRA-2 for biofilms growth does not disrupt the function of the bioreactor array. IMPORTANCE The MBRA-2 with media rails provides an accessible system for investigators to culture heterogenous, suspended biofilms under constant flow. |
| first_indexed | 2024-04-10T15:22:02Z |
| format | Article |
| id | doaj.art-da3b1bdddd4b4c37b261a67f1c35616b |
| institution | Directory Open Access Journal |
| issn | 2165-0497 |
| language | English |
| last_indexed | 2024-04-10T15:22:02Z |
| publishDate | 2023-02-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | Microbiology Spectrum |
| spelling | doaj.art-da3b1bdddd4b4c37b261a67f1c35616b2023-02-14T14:15:49ZengAmerican Society for MicrobiologyMicrobiology Spectrum2165-04972023-02-0111110.1128/spectrum.02928-22MBRA-2: a Modified Chemostat System to Culture BiofilmsJustin N. Jens0Daniel J. Breiner1Rachel L. Neve2Matilda M. Fiebig3Vanessa V. Phelan4Department of Pharmaceutical Sciences, Skaggs School of Pharmacy and Pharmaceutical Sciences, University of Colorado Anschutz Medical Campus, Aurora, Colorado, USADepartment of Pharmaceutical Sciences, Skaggs School of Pharmacy and Pharmaceutical Sciences, University of Colorado Anschutz Medical Campus, Aurora, Colorado, USADepartment of Immunology and Microbiology, School of Medicine, University of Colorado Anschutz Medical Campus, Aurora, Colorado, USADepartment of Pharmaceutical Sciences, Skaggs School of Pharmacy and Pharmaceutical Sciences, University of Colorado Anschutz Medical Campus, Aurora, Colorado, USADepartment of Pharmaceutical Sciences, Skaggs School of Pharmacy and Pharmaceutical Sciences, University of Colorado Anschutz Medical Campus, Aurora, Colorado, USAABSTRACT Culture-dependent approaches for investigating microbial ecology aim to model the nutrient content of specific environments by simplifying the system for high-resolution molecular analysis. These in vitro systems are enticing due to their increased throughput compared to animal models, flexibility in modulating nutrient content and community composition, scaling of culture volume to isolate biological molecules, and control of environmental parameters, such as temperature, humidity, and nutrient flow. However, different devices are used to investigate homogenous, planktonic microbial communities and heterogeneous biofilms. Here, we present the minibioreactor array 2 (MBRA-2) with media rails, a benchtop multireactor system derived from the MBRA system that enables researchers to use the same system to grow planktonic and biofilm cultures. We simplified flow through the system and reduced contamination, leakage, and time required for array assembly by designing and implementing a reusable media rail to replace the branched tubing traditionally used to convey media through chemostat arrays. Additionally, we altered the structure of the six-bioreactor strip to incorporate a removable lid to provide easy access to the bioreactor wells, enabling biofilm recovery and thorough cleaning for reuse. Using Pseudomonas aeruginosa, a model biofilm-producing organism, we show that the technical improvements of the MBRA-2 for biofilms growth does not disrupt the function of the bioreactor array. IMPORTANCE The MBRA-2 with media rails provides an accessible system for investigators to culture heterogenous, suspended biofilms under constant flow.https://journals.asm.org/doi/10.1128/spectrum.02928-22MBRA-2Pseudomonas aeruginosabiofilmschemostat cultures |
| spellingShingle | Justin N. Jens Daniel J. Breiner Rachel L. Neve Matilda M. Fiebig Vanessa V. Phelan MBRA-2: a Modified Chemostat System to Culture Biofilms Microbiology Spectrum MBRA-2 Pseudomonas aeruginosa biofilms chemostat cultures |
| title | MBRA-2: a Modified Chemostat System to Culture Biofilms |
| title_full | MBRA-2: a Modified Chemostat System to Culture Biofilms |
| title_fullStr | MBRA-2: a Modified Chemostat System to Culture Biofilms |
| title_full_unstemmed | MBRA-2: a Modified Chemostat System to Culture Biofilms |
| title_short | MBRA-2: a Modified Chemostat System to Culture Biofilms |
| title_sort | mbra 2 a modified chemostat system to culture biofilms |
| topic | MBRA-2 Pseudomonas aeruginosa biofilms chemostat cultures |
| url | https://journals.asm.org/doi/10.1128/spectrum.02928-22 |
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