Microsporidia infection alters C. elegans lipid levels

Hala Tamim El Jarkass; JiHae Jeon; Nicholas O. Burton; Aaron W. Reinke

doi:10.1371/journal.pone.0327188

Peer Review History

Original SubmissionFebruary 17, 2025
Decision Letter - Paul Dean, Editor Dear Dr. Reinke, Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process. ============================== The reviewers provided positive reviews about the work and only require minor revision. I don't feel additional experimental work is needed. Please address the reviewers' points and submit your revised version. ============================== Please submit your revised manuscript by May 03 2025 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org . 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C. elegans strains were provided by the Caenorhabditis Genetics Center, which is funded by the National Institutes of Health (NIH) Office of Research Infrastructure Programs Grant P40 OD010440 and we thank WormBase. Funding: This work was supported by the Canadian Institutes of Health Research grant no. 400784 (to A. R.) Competing interests: The authors declare that they have no competing interests. Data availability: All data needed to evaluate the conclusions in the paper are present in the paper and supplementary materials. Source Data are provided in Data S1.” We note that you have provided funding information that is currently declared in your Funding Statement. However, funding information should not appear in the Acknowledgments section or other areas of your manuscript. We will only publish funding information present in the Funding Statement section of the online submission form. Please remove any funding-related text from the manuscript and let us know how you would like to update your Funding Statement. Currently, your Funding Statement reads as follows: “This work was supported by the Canadian Institutes of Health Research grant no. 400784 (to A. R.)” Please include your amended statements within your cover letter; we will change the online submission form on your behalf. 5. We notice that your supplementary materials are included in the manuscript file. Please remove them and upload them with the file type 'Supporting Information'. Please ensure that each Supporting Information file has a legend listed in the manuscript after the references list. 6. Please review your reference list to ensure that it is complete and correct. If you have cited papers that have been retracted, please include the rationale for doing so in the manuscript text, or remove these references and replace them with relevant current references. Any changes to the reference list should be mentioned in the rebuttal letter that accompanies your revised manuscript. If you need to cite a retracted article, indicate the article’s retracted status in the References list and also include a citation and full reference for the retraction notice. [Note: HTML markup is below. Please do not edit.] Reviewers' comments: Reviewer's Responses to Questions Comments to the Author 1. Is the manuscript technically sound, and do the data support the conclusions? Reviewer #1: Yes Reviewer #2: Yes ******** 2. Has the statistical analysis been performed appropriately and rigorously? -->?> Reviewer #1: Yes Reviewer #2: Yes ****** 3. Have the authors made all data underlying the findings in their manuscript fully available??> The PLOS Data policy Reviewer #1: Yes Reviewer #2: Yes ****** 4. Is the manuscript presented in an intelligible fashion and written in standard English??> Reviewer #1: Yes Reviewer #2: Yes ****** Reviewer #1: Excellent manuscript, with some novel and interesting information presented. My comments are only minor, and refer mainly to the presentation of statistical output/detail. In several of your figures, you present a series of plots. These are presented really well with individual points represented; however, in one or two cases there is a bar plot with error without points presented (or perhaps they are covered by error?). Please can these be presented as box-plots to allow the reader to gather further visual information about the variance between your replicates, while including the points in a similar way to the other plots. In various places throughout the manuscript, a p-value is listed to support a finding, but the accompanying statistic and base information on data normality are missing. I cannot determine if data normality was checked, supporting statistic use - I don't believe this is specifically noted in the methods. Please can this be made clearer in the methods. In addition, where a p-value is listed, please include the statistic that was used to provide this value. This could also include the degrees of freedom and any accompanying value referring to variance (where relevant). One point for the discussion. I wonder if you might elaborate on what your findings may mean for microsporidian culture. In many cases, lipid content in cell line choice is not a commonly explored parameter. Perhaps this is an important factor to consider for many species? If lipid levels change, this could have an effect on an ability to culture certain species. Reviewer #2: The paper by El Jarkass and colleagues aimed to determine the links between lipidome and microsporidian infections in C. elegans. The work has been very well performed, and represent an important addition to our current knowledge of microsporidian infections and how it affects the host metabolism. I have no major comments, as I found the work to be well executed and timely. One suggestion would be to ensure that the results are placed into the context of the host species - for example, can we expect the lipidome of all or most hosts to be similarly affected, or does the C. elegans genome carry some uniqueness from that perspective?. ****** what does this mean? ). If published, this will include your full peer review and any attached files. If you choose “no”, your identity will remain anonymous but your review may still be made public. Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy Reviewer #1: Yes: Dr Jamie Bojko Reviewer #2: No ******** [NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.] While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/ . PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at figures@plos.org https://doi.org/10.1371/journal.pone.0327188.r001
Revision 1
11 May 2025 Author Response We thank the reviewers for their comments. We have provided a revised manuscript (where all changes are tracked) to address the reviewers’ concerns. Points raised by the reviewers have been incorporated into the manuscript and we have provided a point-by-point response to each of the reviewer’s comments: Reviewer #1: Reviewer #1: Excellent manuscript, with some novel and interesting information presented. My comments are only minor, and refer mainly to the presentation of statistical output/detail. We thank the reviewer for their kind comments. In several of your figures, you present a series of plots. These are presented really well with individual points represented; however, in one or two cases there is a bar plot with error without points presented (or perhaps they are covered by error?). Please can these be presented as box-plots to allow the reader to gather further visual information about the variance between your replicates, while including the points in a similar way to the other plots. Yes, there are several cases where we present normalized bar plots, where it is difficult to see the individual points. We have now changed these plots to just show the points and standard deviation. In once case, figure 2E, the control is normalized to 1, so there are no error bars to be shown in the control samples in this plot. In various places throughout the manuscript, a p-value is listed to support a finding, but the accompanying statistic and base information on data normality are missing. I cannot determine if data normality was checked, supporting statistic use - I don't believe this is specifically noted in the methods. Please can this be made clearer in the methods. In addition, where a p-value is listed, please include the statistic that was used to provide this value. This could also include the degrees of freedom and any accompanying value referring to variance (where relevant). To address this comment, we have added a statistics section to the methods and modified the legends to include the statistical test that was used for each figure panel. We also checked the normality of our data, and this information has been added to the statistics section of the methods. One point for the discussion. I wonder if you might elaborate on what your findings may mean for microsporidian culture. In many cases, lipid content in cell line choice is not a commonly explored parameter. Perhaps this is an important factor to consider for many species? If lipid levels change, this could have an effect on an ability to culture certain species. We have modified the discussion in lines 169 to 175 to mention how fatty acids can affect growth of other parasites in cell culture and microsporidia in several animals, and how this approach may be used to culture microsporidia such as E. bieneusi. Reviewer #2: The paper by El Jarkass and colleagues aimed to determine the links between lipidome and microsporidian infections in C. elegans. The work has been very well performed, and represent an important addition to our current knowledge of microsporidian infections and how it affects the host metabolism. We thank the reviewer for their kind comments. I have no major comments, as I found the work to be well executed and timely. One suggestion would be to ensure that the results are placed into the context of the host species - for example, can we expect the lipidome of all or most hosts to be similarly affected, or does the C. elegans genome carry some uniqueness from that perspective? The strong impact of microsporidia on host lipid metabolism has now been reported in in fruit flies, shrimp, crabs, and silkworms and honey bees. Although there are differences in the exact lipid species being affected, lack of standardized experimental approaches make comparisons difficult and future work will be necessary to determine the importance of these differences. We have modified the discussion in lines 175 to 177 to mention and reference other hosts where changes to lipid metabolism caused by microsporidia have been observed. Attachments Attachment Submitted filename: plosone_response_letter_0511125.pdf https://doi.org/10.1371/journal.pone.0327188.r002
Decision Letter - Paul Dean, Editor Microsporidia infection alters C. elegans lipid levels PONE-D-25-08621R1 Dear Dr. Reinke, We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements. Within one week, you’ll receive an e-mail detailing the required amendments. When these have been addressed, you’ll receive a formal acceptance letter and your manuscript will be scheduled for publication. An invoice will be generated when your article is formally accepted. Please note, if your institution has a publishing partnership with PLOS and your article meets the relevant criteria, all or part of your publication costs will be covered. Please make sure your user information is up-to-date by logging into Editorial Manager at Editorial Manager® and clicking the ‘Update My Information' link at the top of the page. If you have any questions relating to publication charges, please contact our Author Billing department directly at authorbilling@plos.org. If your institution or institutions have a press office, please notify them about your upcoming paper to help maximize its impact. If they’ll be preparing press materials, please inform our press team as soon as possible -- no later than 48 hours after receiving the formal acceptance. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information, please contact onepress@plos.org. Kind regards, Paul Dean Academic Editor PLOS ONE Additional Editor Comments (optional): Thank you for your revised version of the manuscript and for addressing the points of the reviewers. Reviewers' comments: https://doi.org/10.1371/journal.pone.0327188.r003
Formally Accepted
Acceptance Letter - Paul Dean, Editor PONE-D-25-08621R1 PLOS ONE Dear Dr. Reinke, I'm pleased to inform you that your manuscript has been deemed suitable for publication in PLOS ONE. Congratulations! Your manuscript is now being handed over to our production team. At this stage, our production department will prepare your paper for publication. This includes ensuring the following: * All references, tables, and figures are properly cited * All relevant supporting information is included in the manuscript submission, * There are no issues that prevent the paper from being properly typeset You will receive further instructions from the production team, including instructions on how to review your proof when it is ready. Please keep in mind that we are working through a large volume of accepted articles, so please give us a few days to review your paper and let you know the next and final steps. Lastly, if your institution or institutions have a press office, please let them know about your upcoming paper now to help maximize its impact. If they'll be preparing press materials, please inform our press team within the next 48 hours. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information, please contact onepress@plos.org. If we can help with anything else, please email us at customercare@plos.org. Thank you for submitting your work to PLOS ONE and supporting open access. Kind regards, PLOS ONE Editorial Office Staff on behalf of Dr. Paul Dean Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0327188.r004

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