Characterizing nitrogen cycling microorganisms and genes in sediments of the Three Gorges Reservoir

Chang Han; Zhiyuan Chen; Yihui Xiao; Ting Yang; Haoyang Shi; Huiqun Cao; Wenjun Yang; Ping Gong

doi:10.1371/journal.pone.0324051

Peer Review History

Original SubmissionAugust 30, 2024
30 Aug 2024 Author Response https://doi.org/10.1371/journal.pone.0324051.r001
Decision Letter - Farhan Hafeez, Editor Dear Dr. Yang, 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. Please submit your revised manuscript by Nov 30 2024 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 . When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file. A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'. 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Please remove your figures from within your manuscript file, leaving only the individual TIFF/EPS image files, uploaded separately. These will be automatically included in the reviewers’ PDF. Additional Editor Comments: Dear Author(s), I write to you regarding the manuscript PONE-D-24-37897 entitled "Characterizing Nitrogen-Cycling Microorganisms and Genes in Sediments of the Three Gorges Reservoir" which was submitted to PLOS One. The reviewers have now commented on the manuscript. You will see that they are advising substantial revision of the manuscript. In addition to the reviewers’ comments, the overall write-up needs considerable improvement. When revising your work, please submit a list of changes or a rebuttal against each point being raised through track changes mode or by using bold or colored text. Please revise the manuscript strictly according to the appended comments from the reviewers. While revising, need is to double check that all the references cited within the text have corresponding references. Sincerely, [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: No ****** Reviewer #1: The manuscript ‘Characterizing Nitrogen-Cycling Microorganisms and Genes in Sediments of the Three Gorges Reservoir’ focuses an important topic of investigating microbial-driven nitrogen cycle processes in the sediments of the Three Gorges Reservoir and assess the overall state of nitrogen cycling. The current manuscript comes in the domain of the journal and I appreciate the authors for their research work. However, due to various issues across different sections of manuscript, it needs revision. Overall, sentence structure needs to be improved. Some of general comments/questions are listed below while more specific questions and comments are inserted in the attached PDF file. In abstract elaborate briefly, what authors want to say when write ‘46 types of nitrogen conversion functional genes’, nitrogen conversion not accurate there. Further. it’s not ‘refine’ rather this study aims to investigate or report. Please rephrase. The current structure of the introduction is not well organized. Additionally, the last part of the introduction needs to be revised, considering the main theme/objectives and findings of the study. Materials and Methods are very descriptive, it’s more like a thesis document especially in methods section, there are a large number of sub-headings in methods. And the representation of data also needs to be extensively improved. Further, the figures could be merged, or only key data may be included instead of having huge numbers which could easily be merged. The results are explained while the Discussion section could be improved, for example in addition to discussing the results in line with previous reports, key findings need to be discussed and highlighted. The section 4 overall, and specifically with focus on ‘ammonia oxidation requires specific environmental conditions’ is also like a thesis document, much detail and redundant rather than just discussing the key findings. Another very crucial aspect of the work i.e. how the ‘variations in sediment particle size’ influences the N cycling is described but lacks in depth analyses, for example, the relative abundance of these functional guilds across various environments could also be compared to enlighten the current results. Again, like other sections of the manuscript, Discussion, and Conclusions sections also needs to be comprehensive (instead of detailed paras and redundancy at places), the focus could only be on the key findings of the work. Reviewer #2: The research work described the Characterization of Nitrogen-Cycling Microorganisms and Genes in Sediments. The article reports interesting results, however write up of the article could be a lot better than this draft. Language of introduction could be improved. There are so many sub-sections in the methodology section which do not follow the article style. A huge number of figures are also not desired. The quality of Figure 2a is not readable. Nitrogen is a very crucial element across sediments, discussion section needs to be extensively improved considering the nitrogen importance across sediments. ****** 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: No 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 . Please note that Supporting Information files do not need this step. Attachments Attachment Submitted filename: PONE-D-24-37897 (1).pdf https://doi.org/10.1371/journal.pone.0324051.r002
Revision 1
23 Dec 2024 Author Response Reviewer #1: The manuscript ‘Characterizing Nitrogen-Cycling Microorganisms and Genes in Sediments of the Three Gorges Reservoir’ focuses an important topic of investigating microbial-driven nitrogen cycle processes in the sediments of the Three Gorges Reservoir and assess the overall state of nitrogen cycling. The current manuscript comes in the domain of the journal and I appreciate the authors for their research work. However, due to various issues across different sections of manuscript, it needs revision. Overall, sentence structure needs to be improved. Some of general comments/questions are listed below while more specific questions and comments are inserted in the attached PDF file. In abstract elaborate briefly, what authors want to say when write ‘46 types of nitrogen conversion functional genes’, nitrogen conversion not accurate there. Further. it’s not ‘refine’ rather this study aims to investigate or report. Please rephrase. The current structure of the introduction is not well organized. Additionally, the last part of the introduction needs to be revised, considering the main theme/objectives and findings of the study. Materials and Methods are very descriptive, it’s more like a thesis document especially in methods section, there are a large number of sub-headings in methods. And the representation of data also needs to be extensively improved. Further, the figures could be merged, or only key data may be included instead of having huge numbers which could easily be merged. The results are explained while the Discussion section could be improved, for example in addition to discussing the results in line with previous reports, key findings need to be discussed and highlighted. The section 4 overall, and specifically with focus on ‘ammonia oxidation requires specific environmental conditions’ is also like a thesis document, much detail and redundant rather than just discussing the key findings. Another very crucial aspect of the work i.e. how the ‘variations in sediment particle size’ influences the N cycling is described but lacks in depth analyses, for example, the relative abundance of these functional guilds across various environments could also be compared to enlighten the current results. Again, like other sections of the manuscript, Discussion, and Conclusions sections also needs to be comprehensive (instead of detailed paras and redundancy at places), the focus could only be on the key findings of the work. Reply: I have already made modifications to all the problematic areas listed in the PDF file according to the comments. I have rewritten the abstract section, reorganizing the language. The sentence "46 types of nitrogen conversion functional genes" has been removed, and the structure of the sentences has been reorganized to ensure accurate expression. I have revised the last part of the introduction, incorporating the research theme/objectives and the investigation results. I have simplified the descriptions in the "Materials and Methods" section and consolidated some overly detailed sub-headings to reduce the complexity of the methods part and ensure that each section is directly relevant and contributes to the research findings. I have eliminated certain analyses that bear relatively low relevance to the overall conclusion and opted to present solely the key data. By doing so, I aim to ensure the effective conveyance of the information's essence while circumventing excessive and redundant data. I have endeavored to render the data presentation more concise and influential, thus conforming to the standards expected of high-quality research papers. While reviewing the manuscript, I noticed in the section “3.5 Correlative analysis of environmental factors” that some data for the environmental factor had been incorrectly entered during the initial analysis and graphing. I have since input the correct data, conducted a new analysis and redrafted the figures, and included the updated results in the manuscript. Similarly, due to some incorrect inputs of environmental factors initially, there were also issues with the analysis in the section "3.6 Random forest analysis". Therefore, after inputting the correct data, I conducted a new analysis and redrafted the figures, and have included the updated results in the manuscript. I have made improvements to the discussion section, focusing on and emphasizing the key findings. I have eliminated redundant parts and conducted an in-depth analysis of how “variations in sediment particle size” influences the nitrogen cycle. The discussion and conclusion sections have been thoroughly revised to highlight the critical findings of this study. Reviewer #2: The research work described the Characterization of Nitrogen-Cycling Microorganisms and Genes in Sediments. The article reports interesting results, however write up of the article could be a lot better than this draft. Language of introduction could be improved. There are so many sub-sections in the methodology section which do not follow the article style. A huge number of figures are also not desired. The quality of Figure 2a is not readable. Nitrogen is a very crucial element across sediments, discussion section needs to be extensively improved considering the nitrogen importance across sediments. Reply: I have improved the language in the introduction section. The methods section has been revised by removing unnecessary subsections to streamline the presentation and improve clarity. A significant amount of analysis and figures with weak relevance to this study have been removed. I have enhanced the quality of the figures and included all high-definition figures in the appendix. The discussion section has been thoroughly modified. Attachments Attachment Submitted filename: Response to Reviewers.docx https://doi.org/10.1371/journal.pone.0324051.r003
Decision Letter - Farhan Hafeez, Editor Dear Dr. Yang, 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. Please submit your revised manuscript by Feb 10 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 . When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file. A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'. A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'. An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'. If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter. If applicable, we recommend that you deposit your laboratory protocols in protocols.io to enhance the reproducibility of your results. Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: https://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols . Additionally, PLOS ONE offers an option for publishing peer-reviewed Lab Protocol articles, which describe protocols hosted on protocols.io. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols . We look forward to receiving your revised manuscript. Kind regards, Farhan Hafeez, Ph.D. Academic Editor PLOS ONE Additional Editor Comments: Dear Authors, The revised version of the manuscript has been reviewed. Thanks to the authors for taking into account the previous comments. However, there are still the comments to be addressed as in appended text. The important comment: “Further, the figures could be merged, or only key data may be included instead of having huge numbers which could easily be merged” yet needs to be considered. Yes, the authors have just combined the figures, rather there is need to merge the figures for example in the form of panel. Currently, data presentation is more like a thesis document. Sincerely, [Note: HTML markup is below. Please do not edit.] [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 . Please note that Supporting Information files do not need this step. https://doi.org/10.1371/journal.pone.0324051.r004
Revision 2
18 Feb 2025 Author Response Reviewer #1: The manuscript ‘Characterizing Nitrogen-Cycling Microorganisms and Genes in Sediments of the Three Gorges Reservoir’ focuses an important topic of investigating microbial-driven nitrogen cycle processes in the sediments of the Three Gorges Reservoir and assess the overall state of nitrogen cycling. The current manuscript comes in the domain of the journal and I appreciate the authors for their research work. However, due to various issues across different sections of manuscript, it needs revision. Overall, sentence structure needs to be improved. Some of general comments/questions are listed below while more specific questions and comments are inserted in the attached PDF file. In abstract elaborate briefly, what authors want to say when write ‘46 types of nitrogen conversion functional genes’, nitrogen conversion not accurate there. Further. it’s not ‘refine’ rather this study aims to investigate or report. Please rephrase. The current structure of the introduction is not well organized. Additionally, the last part of the introduction needs to be revised, considering the main theme/objectives and findings of the study. Materials and Methods are very descriptive, it’s more like a thesis document especially in methods section, there are a large number of sub-headings in methods. And the representation of data also needs to be extensively improved. Further, the figures could be merged, or only key data may be included instead of having huge numbers which could easily be merged. The results are explained while the Discussion section could be improved, for example in addition to discussing the results in line with previous reports, key findings need to be discussed and highlighted. The section 4 overall, and specifically with focus on ‘ammonia oxidation requires specific environmental conditions’ is also like a thesis document, much detail and redundant rather than just discussing the key findings. Another very crucial aspect of the work i.e. how the ‘variations in sediment particle size’ influences the N cycling is described but lacks in depth analyses, for example, the relative abundance of these functional guilds across various environments could also be compared to enlighten the current results. Again, like other sections of the manuscript, Discussion, and Conclusions sections also needs to be comprehensive (instead of detailed paras and redundancy at places), the focus could only be on the key findings of the work. Reply: I have already made modifications to all the problematic areas listed in the PDF file according to the comments. I have rewritten the abstract section, reorganizing the language. The sentence "46 types of nitrogen conversion functional genes" has been removed, and the structure of the sentences has been reorganized to ensure accurate expression. I have revised the last part of the introduction, incorporating the research theme/objectives and the investigation results. I have simplified the descriptions in the "Materials and Methods" section and consolidated some overly detailed sub-headings to reduce the complexity of the methods part and ensure that each section is directly relevant and contributes to the research findings. I have eliminated certain analyses that bear relatively low relevance to the overall conclusion and opted to present solely the key data. By doing so, I aim to ensure the effective conveyance of the information's essence while circumventing excessive and redundant data. I have endeavored to render the data presentation more concise and influential, thus conforming to the standards expected of high-quality research papers. While reviewing the manuscript, I noticed in the section “3.5 Correlative analysis of environmental factors” that some data for the environmental factor had been incorrectly entered during the initial analysis and graphing. I have since input the correct data, conducted a new analysis and redrafted the figures, and included the updated results in the manuscript. Similarly, due to some incorrect inputs of environmental factors initially, there were also issues with the analysis in the section "3.6 Random forest analysis". Therefore, after inputting the correct data, I conducted a new analysis and redrafted the figures, and have included the updated results in the manuscript. I have made improvements to the discussion section, focusing on and emphasizing the key findings. I have eliminated redundant parts and conducted an in-depth analysis of how “variations in sediment particle size” influences the nitrogen cycle. The discussion and conclusion sections have been thoroughly revised to highlight the critical findings of this study. Reviewer #2: The research work described the Characterization of Nitrogen-Cycling Microorganisms and Genes in Sediments. The article reports interesting results, however write up of the article could be a lot better than this draft. Language of introduction could be improved. There are so many sub-sections in the methodology section which do not follow the article style. A huge number of figures are also not desired. The quality of Figure 2a is not readable. Nitrogen is a very crucial element across sediments, discussion section needs to be extensively improved considering the nitrogen importance across sediments. Reply: I have improved the language in the introduction section. The methods section has been revised by removing unnecessary subsections to streamline the presentation and improve clarity. A significant amount of analysis and figures with weak relevance to this study have been removed. I have enhanced the quality of the figures and included all high-definition figures in the appendix. The discussion section has been thoroughly modified. Attachments Attachment Submitted filename: Response_to_Reviewers_auresp_2.docx https://doi.org/10.1371/journal.pone.0324051.r005
Decision Letter - Farhan Hafeez, Editor Dear Dr. Yang, 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. Please submit your revised manuscript by Apr 13 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 . When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file. A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'. A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'. An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'. If applicable, we recommend that you deposit your laboratory protocols in protocols.io to enhance the reproducibility of your results. Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: https://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols . Additionally, PLOS ONE offers an option for publishing peer-reviewed Lab Protocol articles, which describe protocols hosted on protocols.io. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols . We look forward to receiving your revised manuscript. Kind regards, Farhan Hafeez, Ph.D. Academic Editor PLOS ONE Journal Requirements: 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. Additional Editor Comments: Dear Authors, The revised version (R2) of the manuscript has been reviewed. I wonder, the authors just removed a few figures, while the comment was “Further, the figures could be merged, or only key data may be included instead of having huge numbers which could easily be merged”. For example, 3c, 3d, 5a and 6a are excluded. Either these were not important anymore or could these be merged (merged – means the data, the raw data could be combined to have composite figures). Currently, data presentation is more like a thesis document. Sincerely, [Note: HTML markup is below. Please do not edit.] [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 . Please note that Supporting Information files do not need this step. https://doi.org/10.1371/journal.pone.0324051.r006
Revision 3
15 Apr 2025 Author Response Journal Requirements: 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. Reply: I have checked the reference list to ensure it is complete and accurate. Additional Editor Comments: Dear Authors, The revised version (R2) of the manuscript has been reviewed. I wonder, the authors just removed a few figures, while the comment was “Further, the figures could be merged, or only key data may be included instead of having huge numbers which could easily be merged”. For example, 3c, 3d, 5a and 6a are excluded. Either these were not important anymore or could these be merged (merged – means the data, the raw data could be combined to have composite figures). Currently, data presentation is more like a thesis document. Reply: Figures 3c, 3d, 5a, and 6a have been deleted because they are no longer important. Attachments Attachment Submitted filename: Response_to_Reviewers_auresp_3.docx https://doi.org/10.1371/journal.pone.0324051.r007
Decision Letter - Farhan Hafeez, Editor Characterizing nitrogen cycling microorganisms and genes in sediments of the Three Gorges Reservoir PONE-D-24-37897R3 Dear Dr. Han, 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, Farhan Hafeez, Ph.D. Academic Editor PLOS ONE Additional Editor Comments (optional): Dear Author(s), I write to you regarding the manuscript PONE-D-24-37897R3 entitled "Characterizing Nitrogen-Cycling Microorganisms and Genes in Sediments of the Three Gorges Reservoir" which was submitted to PLOS One. Though I still wonder with the reply of the authors that ‘Figures 3c, 3d, 5a, and 6a have been deleted because they are no longer important’ in response to my comment ‘Either these were not important anymore or could these be merged’. However, most of the comments are addressed, hence the manuscript now carries the required potential. I am pleased to inform you that your manuscript is now sufficiently improved for possible publication in PLOS One. The formal acceptance is subject to fulfillment of all the technical requirements. On behalf of PLOS One, I appreciate you for your contribution. Please keep us in mind for any future work that you consider to be appropriate for our readers. Sincerely, Farhan Hafeez, PhD Reviewers' comments: https://doi.org/10.1371/journal.pone.0324051.r008
Formally Accepted
Acceptance Letter - Farhan Hafeez, Editor PONE-D-24-37897R3 PLOS ONE Dear Dr. Yang, 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. Farhan Hafeez Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0324051.r009

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