Presentation Information
[P03-380]Novel screening method for environmental pollutant-degrading bacteria via bacteriophage-mediated selective depletion
○Madoka Kashiki1, Hiroaki Iwaki2, Kenji Okano2 (1. Kansai University Graduated School (Japan), 2. Kansai University (Japan))
Keywords:
bacteriophage,pollutant-degrading bacteria
[Purpose]
Environmental pollution caused by the release of chemicals into the environment is a serious global issue. In recent years, in situ bioremediation has emerged a promising approach for sustainable environmental cleanup. To enable effective bioremediation, extensive efforts have been made to explore and utilize diverse bacteria capable of efficiently degrading target pollutants. However, conventional isolation methods are often dominated by fast-growing bacteria under given culture conditions. This cultivation bias makes it difficult to isolate rare and slow-growing bacteria, leading many potentially useful bacteria undiscovered. To overcome this limitation, selective removal of dominant species from microbial communities is an effective strategy.Our laboratory has developed methods for species-specific removal of target bacteria from microbial communities. This approach exploits the host-specific lytic activity of bacteriophages to eliminate dominant species. We anticipate that applying this strategy to enrichment cultures will enable the isolation of uncultured or overlooked bacteria that are otherwise outcompeted under standard culture conditions.
[Results]
In this study, we evaluated the feasibility of eliminating dominant species with bacteriophages in a model soil microbiota. First, we isolated 2-nitrobenzoate (2-NBA)-degrading bacteria from soil. Following enrichment, four 2-NBA-degrading bacteria species; Cupriavidus oxalaticus, Cupriavidus necator, Ensifer adhaerens, and Achromobacter anxifer were isolated. Next, we attempted to isolate bacteriophages infecting these isolates from sewage samples. As a result, a phage infecting C. oxalaticus was successfully isolated and designated as ΦCoMK1.As an initial proof-of-concept for bacteriophage-mediated removal of dominant species from environmental samples, we constructed a model soil microbiota consisting of four isolates, in which C. oxalaticus was dominant. Upon addition of ΦCoMK1 and cultivation for 48 hours, the relative abundance of C. oxalaticus decreased from 99.9% to 0.4%, accompanied by increased proliferation of the other three species.
[Conclusion]
These results demonstrate that phage addition enables the elimination of dominant species, with a corresponding increase in minor bacterial population within microbial communities. This approach would provide an effective means to access previously underexplored microbial resources. As a prospect, we plan to apply this method to natural soil microbiota to isolate novel pollutant-degrading bacteria.
Environmental pollution caused by the release of chemicals into the environment is a serious global issue. In recent years, in situ bioremediation has emerged a promising approach for sustainable environmental cleanup. To enable effective bioremediation, extensive efforts have been made to explore and utilize diverse bacteria capable of efficiently degrading target pollutants. However, conventional isolation methods are often dominated by fast-growing bacteria under given culture conditions. This cultivation bias makes it difficult to isolate rare and slow-growing bacteria, leading many potentially useful bacteria undiscovered. To overcome this limitation, selective removal of dominant species from microbial communities is an effective strategy.Our laboratory has developed methods for species-specific removal of target bacteria from microbial communities. This approach exploits the host-specific lytic activity of bacteriophages to eliminate dominant species. We anticipate that applying this strategy to enrichment cultures will enable the isolation of uncultured or overlooked bacteria that are otherwise outcompeted under standard culture conditions.
[Results]
In this study, we evaluated the feasibility of eliminating dominant species with bacteriophages in a model soil microbiota. First, we isolated 2-nitrobenzoate (2-NBA)-degrading bacteria from soil. Following enrichment, four 2-NBA-degrading bacteria species; Cupriavidus oxalaticus, Cupriavidus necator, Ensifer adhaerens, and Achromobacter anxifer were isolated. Next, we attempted to isolate bacteriophages infecting these isolates from sewage samples. As a result, a phage infecting C. oxalaticus was successfully isolated and designated as ΦCoMK1.As an initial proof-of-concept for bacteriophage-mediated removal of dominant species from environmental samples, we constructed a model soil microbiota consisting of four isolates, in which C. oxalaticus was dominant. Upon addition of ΦCoMK1 and cultivation for 48 hours, the relative abundance of C. oxalaticus decreased from 99.9% to 0.4%, accompanied by increased proliferation of the other three species.
[Conclusion]
These results demonstrate that phage addition enables the elimination of dominant species, with a corresponding increase in minor bacterial population within microbial communities. This approach would provide an effective means to access previously underexplored microbial resources. As a prospect, we plan to apply this method to natural soil microbiota to isolate novel pollutant-degrading bacteria.
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