The Microbiome of C. elegans Enhances Growth of S. aureus During Infection
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Staphylococcus aureus is a significant human pathogen associated with a wide range of infections. The microbiome, consisting of diverse microbial communities, has been recognized as a critical factor in modulating host-pathogen interactions. This study aimed to investigate the role of the Caenorhabditis elegans microbiome in Staphylococcus aureus infection by transferring worms from the standard laboratory food source (OP50) or a microbiome sample to S. aureus cultures. Our findings revealed that the C. elegans microbiome paradoxically enhanced Staphylococcus aureus colonization, resulting in increased pathogen burden compared to worms transferred from E.coli OP50. These results underscore the importance of the microbiome in modulating host susceptibility.
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Please be sure to address all reviewer comments and include any additional information that is requested in the revised manuscript. If you object to the proposed additional experiments, sufficient justification will be needed. Additionally, it's unclear why the manuscript has sections for an impact statement and outcome that have no actual content in them. Please adjust these sections accordingly.
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Comments to Author
The manuscript presents intriguing findings. I have few queries especially in the methodology section. I request the authors to clarify the same. Lines 114-116: Is the optimal temperature for both strains 37 °C? What is the media recommended by the culture collection center for culturing the microbiome strains? Please provide the rationale for choosing 25 °C for culturing the microbiome strains Line 117: I request the authors to provide more details about 'The microbiome mix' Authors may include a separate section on 'Statistical Analysis' in the methodology I request that the authors follow a consistent format throughout the manuscript, specifically regarding the depiction of bacterial names. In some sections, both the bacterial name and strain number are provided, while in others, only the strain …
Comments to Author
The manuscript presents intriguing findings. I have few queries especially in the methodology section. I request the authors to clarify the same. Lines 114-116: Is the optimal temperature for both strains 37 °C? What is the media recommended by the culture collection center for culturing the microbiome strains? Please provide the rationale for choosing 25 °C for culturing the microbiome strains Line 117: I request the authors to provide more details about 'The microbiome mix' Authors may include a separate section on 'Statistical Analysis' in the methodology I request that the authors follow a consistent format throughout the manuscript, specifically regarding the depiction of bacterial names. In some sections, both the bacterial name and strain number are provided, while in others, only the strain number is included. A uniform approach should be maintained throughout. Additionally, I request the authors to provide the full form of any abbreviations the first time they appear in the text Line 147: Why Mannitol Salt Agar medium was used at this juncture? Figures referenced should be briefly discussed in the results section The discussion section may be further strengthened A thorough proofreading for grammatical consistency and clarity is essential to enhance the manuscript's overall readability
Please rate the manuscript for methodological rigour
Good
Please rate the quality of the presentation and structure of the manuscript
Good
To what extent are the conclusions supported by the data?
Strongly support
Do you have any concerns of possible image manipulation, plagiarism or any other unethical practices?
No
Is there a potential financial or other conflict of interest between yourself and the author(s)?
No
If this manuscript involves human and/or animal work, have the subjects been treated in an ethical manner and the authors complied with the appropriate guidelines?
Yes
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Comments to Author
C. elegans is an excellent model for studying bacterial colonization and the ecological factors affecting colonization success. I'm pleased you chose C. elegans to investigate the impact of S. aureus colonization on the host. Including a mock C. elegans microbiome community is also a compelling aspect of your study. However, I have identified several issues in your manuscript that need attention. Firstly, the observed increase in S. aureus abundance may be due to either its outcompetition of the mock community or inadequate washing of the worms before sequencing. The methods section does not address this adequately. If the worms are grown on S. aureus, they might retain significant amounts of bacteria during washing. I recommend repeating the experiment with an improved washing protocol: transfer the …
Comments to Author
C. elegans is an excellent model for studying bacterial colonization and the ecological factors affecting colonization success. I'm pleased you chose C. elegans to investigate the impact of S. aureus colonization on the host. Including a mock C. elegans microbiome community is also a compelling aspect of your study. However, I have identified several issues in your manuscript that need attention. Firstly, the observed increase in S. aureus abundance may be due to either its outcompetition of the mock community or inadequate washing of the worms before sequencing. The methods section does not address this adequately. If the worms are grown on S. aureus, they might retain significant amounts of bacteria during washing. I recommend repeating the experiment with an improved washing protocol: transfer the worms to a media plate (such as Mannitol Salt Agar) without a bacterial lawn and wash them several times. Monitor the empty plate after washing to ensure no residual bacterial growth. Inadequate washing could lead to an overestimation of S. aureus in your sequencing data, which would not accurately reflect colonization. Additionally, consider using reporter isolates to confirm intestinal colonization. Regarding your sample size, did you sequence just three worms? I suggest using three replicate populations of worms, grown under the same conditions on the same day, and lysing them before sequencing. Performing colony counts on different culture media, at least for S. aureus and OP50, would provide further validation. While your use of spent media to assess S. aureus interactions with the mock community bacteria in vitro is a good approach for studying contact-dependent interactions, did you also perform a co-culture assay? A co-culture assay might be more relevant to your in vivo findings. Lastly, the presentation of your data needs improvement. Use survival curves and avoid setting the Y-axis to 150%. By addressing these issues, your study will provide clearer and more reliable insights into the colonization dynamics of S. aureus in C. elegans.
Please rate the manuscript for methodological rigour
Poor
Please rate the quality of the presentation and structure of the manuscript
Very poor
To what extent are the conclusions supported by the data?
Not at all
Do you have any concerns of possible image manipulation, plagiarism or any other unethical practices?
No
Is there a potential financial or other conflict of interest between yourself and the author(s)?
No
If this manuscript involves human and/or animal work, have the subjects been treated in an ethical manner and the authors complied with the appropriate guidelines?
Yes
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