Intra-horn insemination in the alpaca Vicugna pacos: Copulatory wounding and deep sperm deposition

Patricia L. R. Brennan; Stephen Purdy; Sarah J. Bacon

doi:10.1371/journal.pone.0295882

Peer Review History

Original SubmissionNovember 30, 2023
18 Jan 2024 Decision Letter - Jayonta Bhattacharjee, Editor PONE-D-23-40086Intra-horn insemination in the alpaca Vicugna pacos: Copulatory wounding and deep sperm depositionPLOS ONE Dear Dr. Bacon, 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 Mar 03 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. Please include the following items when submitting your revised manuscript: 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, Jayonta Bhattacharjee Academic Editor PLOS ONE Journal Requirements: When submitting your revision, we need you to address these additional requirements. 1. Please ensure that your manuscript meets PLOS ONE's style requirements, including those for file naming. The PLOS ONE style templates can be found at https://journals.plos.org/plosone/s/file?id=wjVg/PLOSOne_formatting_sample_main_body.pdf and https://journals.plos.org/plosone/s/file?id=ba62/PLOSOne_formatting_sample_title_authors_affiliations.pdf 2. Note from Emily Chenette, Editor in Chief of PLOS ONE, and Iain Hrynaszkiewicz, Director of Open Research Solutions at PLOS: Did you know that depositing data in a repository is associated with up to a 25% citation advantage (https://doi.org/10.1371/journal.pone.0230416)? If you’ve not already done so, consider depositing your raw data in a repository to ensure your work is read, appreciated and cited by the largest possible audience. You’ll also earn an Accessible Data icon on your published paper if you deposit your data in any participating repository (https://plos.org/open-science/open-data/#accessible-data). 3. We note that the grant information you provided in the ‘Funding Information’ and ‘Financial Disclosure’ sections do not match. When you resubmit, please ensure that you provide the correct grant numbers for the awards you received for your study in the ‘Funding Information’ section. 4. Thank you for stating the following financial disclosure: "Mount Holyoke College faculty research grant to PLRB. NSF CAREER grant to PLRB, IOS:2042260" Please state what role the funders took in the study. If the funders had no role, please state: ""The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."" If this statement is not correct you must amend it as needed. Please include this amended Role of Funder statement in your cover letter; we will change the online submission form on your behalf. 5. Please include your full ethics statement in the ‘Methods’ section of your manuscript file. In your statement, please include the full name of the IRB or ethics committee who approved or waived your study, as well as whether or not you obtained informed written or verbal consent. If consent was waived for your study, please include this information in your statement as well. 6. Please include captions for your Supporting Information files at the end of your manuscript, and update any in-text citations to match accordingly. Please see our Supporting Information guidelines for more information: http://journals.plos.org/plosone/s/supporting-information. 7. 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? The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented. 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 requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified. Reviewer #1: Yes Reviewer #2: Yes ****** 4. Is the manuscript presented in an intelligible fashion and written in standard English? PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here. Reviewer #1: Yes Reviewer #2: Yes ****** 5. Review Comments to the Author Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters) Reviewer #1: Review of Intra-horn insemination in the alpaca Vicugna pacos: Copulatory wounding and deep sperm deposition. This is an interesting manuscript that has novel data on the process of copulation and sperm transfer in the alpaca. The manuscript is mostly descriptive, but it is very detailed and interesting to read. The results have implications for the evolution of intromission behaviour, the evolution of female and male gonads and sexual conflict theory. I have only some minor comments. Given the wide audience of Plos one, the introduction would greatly benefit from a diagram of the alpaca reproductive system. Materials and methods Local farm where? Examined and photographed immediately after euthanized? It may help other researchers to know what problems were faced with sample preservation of slides, did the stain not work? That is important information so other researchers don’t make the same mistake. Three values after the 0 for p values is more than enough. I suggest values for Table 1 should be a graph, with averages or percentages, making data easier to visualize. Discussion, “female cooperation and receptivity are required for copulation to succeed because copulation is prolonged”not quite following the argument as to why female cooperation is necessary because copulation is long.. In insects copulations can be very lengthy, much more than the 20 min listed here for alpacas, but there still can be sexual conflict if the male is coercing or manipulating the female into mating with him… So I would suggest perhaps limiting this discussion to the ways in which females need to adapt certain positions to allow intromission. Also, is there any evidence for females controlling copula duration as in tephritid flies? Reviewer #2: This is an interesting study of the alpaca natural insemination, there are some mistakes and clarifications and I included my comments in the article PDF attached with this format, for this publication, it is important to know how the animals were cared for this study, the ethic component. ****** 6. PLOS authors have the option to publish the peer review history of their article (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: Diana Perez-Staples Reviewer #2: Yes: Martha Valdivia ******** [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-23-40086 review MV.pdf https://doi.org/10.1371/journal.pone.0295882.r001
Revision 1
23 Feb 2024 Author Response Response to reviewers Brennan, Purdy and Bacon Re: Resubmission of MS PONE-D-23-40086 The reviewers had very minimal comments and we have addressed all of them below. R1: Given the wide audience of Plos one, the introduction would greatly benefit from a diagram of the alpaca reproductive system. AU: We already have a labeled photo in Fig 1, therefore we did not add a diagram. R1: Local farm where? AU: Connecticut. Added R1: Examined and photographed immediately after euthanized? AU: Yes, added R1: It may help other researchers to know what problems were faced with sample preservation of slides, did the stain not work? That is important information so other researchers don’t make the same mistake. AU: We realized that these slides were missing because on the females mated 1hr before euthanasia we had not started yet to use the diff quick stain that allowed us to see immune cells but were using eosin nigrosin stain for live/dead sperm. We have deleted the sentence and instead rewrote this paragraph in the manuscript as follows: “We then used a glass slide to touch each region of the inside of the reproductive tract and stained these slides either with a modified Giemsa stain (Differential Quik Stain Kit, VWR, Radnor PA) to look for the presence of inflammatory cells (unmated and mated +24 hrs) and sperm (mated +24 hrs), or with Semen Morphology Stain (eosin/nigrosin) (mated +1hr)”. R1: Three values after the 0 for p values is more than enough. AU: Fixed R1: I suggest values for Table 1 should be a graph, with averages or percentages, making data easier to visualize. AU: Added Figure 2. Changed the figure captions for all remaining figures R1: Discussion, “female cooperation and receptivity are required for copulation to succeed because copulation is prolonged”not quite following the argument as to why female cooperation is necessary because copulation is long.. In insects copulations can be very lengthy, much more than the 20 min listed here for alpacas, but there still can be sexual conflict if the male is coercing or manipulating the female into mating with him… So I would suggest perhaps limiting this discussion to the ways in which females need to adapt certain positions to allow intromission. Also, is there any evidence for females controlling copula duration as in tephritid flies? AU: We modified this paragraph, now it reads as follows: “However, it seems unlikely that alpacas experience the usual selective pressures that may induce copulatory wounding as a result of sexual conflict; females typically do not mate with multiple males because a single dominant male controls reproductive access to the female group, and female cooperation and receptivity are required for copulation to succeed because females must adopt a prone position to allow intromission, and the lengthy copulation would allow females plenty of opportunity to reject the male by changing position. Either males or females may terminate the copulation, but usually the male stops anywhere between 5-60 minutes after intromission, by standing up. Other times the female may push up or roll to her side and terminate copulation”. Reviewer #2: This is an interesting study of the alpaca natural insemination, there are some mistakes and clarifications and I included my comments in the article PDF attached with this format, for this publication, it is important to know how the animals were cared for this study, the ethic component. Reviewer 2 had 8 spelling and formatting corrections and we have fixed all of them. Since R1 also had a question about housing of the alpaca we added the following statement: “We obtained whole reproductive tracts from adult female alpacas that were being culled for meat processing at a farm in Connecticut, USA, where animals are maintained in accordance with State and federal guidelines concerning livestock management in small farms” Attachments Attachment Submitted filename: Response to reviewers Brennan et al.docx https://doi.org/10.1371/journal.pone.0295882.r002
17 Mar 2024 Decision Letter - Jayonta Bhattacharjee, Editor PONE-D-23-40086R1Intra-horn insemination in the alpaca Vicugna pacos: Copulatory wounding and deep sperm depositionPLOS ONE Dear Dr. Brennan, 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 May 01 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. Please include the following items when submitting your revised manuscript: 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, Jayonta Bhattacharjee 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: Thank you very much for revising the article. I would like to request the authors to address a few minor things. Such as - Page 8, Line 158: particular (F042, 29 and 23% respectively). Where are those data, and what is F042. It would be helpful for the readers if they could track and find that information somewhere in the manuscript or in the additional information files. On the figures, such as in Figures 1 and 2, state that mated, mated day, and unmated. Figure 3 states, mated, mated 24hrs, and unmated. Figure 4 and 5: receptive unmated, mated+1hour, mated+24hour. It would be better to make it uniform style. [Note: HTML markup is below. Please do not edit.] Reviewers' comments: Reviewer's Responses to Questions Comments to the Author 1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation. Reviewer #1: All comments have been addressed Reviewer #2: All comments have been addressed ******** 2. Is the manuscript technically sound, and do the data support the conclusions? The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented. Reviewer #1: Yes Reviewer #2: Yes ****** 3. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: Yes Reviewer #2: Yes ****** 4. Have the authors made all data underlying the findings in their manuscript fully available? The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified. Reviewer #1: Yes Reviewer #2: Yes ****** 5. Is the manuscript presented in an intelligible fashion and written in standard English? PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here. Reviewer #1: Yes Reviewer #2: Yes ****** 6. Review Comments to the Author Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters) Reviewer #1: The authors have adequately replied to the queries. I have no further observations, we need more studies like this one on the female reproductive tract. Reviewer #2: Great and interesting research, your publication has improved in good condition and level. Your results showed the important of type of intercourse in alpacas, the low number of sperm in female tract after 24 hours is unbelievable Congrats!!! ****** 7. PLOS authors have the option to publish the peer review history of their article (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: Diana Pérez-Staples Reviewer #2: Yes: Martha Valdivia ******** [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.0295882.r003
Revision 2
19 Mar 2024 Author Response The editor requested we change the captions in our figures so they would match and we made the changes. Attachments Attachment Submitted filename: Response to reviewers Brennan et al.docx https://doi.org/10.1371/journal.pone.0295882.r004
27 Mar 2024 Decision Letter - Jayonta Bhattacharjee, Editor PONE-D-23-40086R2Intra-horn insemination in the alpaca Vicugna pacos: Copulatory wounding and deep sperm depositionPLOS ONE Dear Dr. Brennan, 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 May 11 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. Please include the following items when submitting your revised manuscript: 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, Jayonta Bhattacharjee 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: Thank you very much for the revised version. I do not know if I am making any mistake, still, I feel a discrepancy between the text and the supplied supplementary table 1. In lines: 152-154: Author mentioned “percentage of bloody pixels (up to 24% for both), but the cervix and uterus also had high percentages in one female in particular (Female 042:, 29 and 23% respectively, data in S1 Table)”. When we go to the S1 table, we see the last row of the supplementary table shows, “mated +24hrs A042 0.06 0 76.19 60.35 13.51 15.47” Is it the same data the author meant, then where are those percentages. May be uterus (left and right horn) is 29% (13.51+15.47=28.98), and the cervix is 76%. Is it true? I am sorry for the misunderstanding. It will be better for the reader to see an easy-understanding table. If there is any discrepancy, I would request the author to put the correct information. [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.0295882.r005
Revision 3
28 Mar 2024 Author Response Comments to the editor We apologize for the confusion. The data in the table are correct, and we revised the text of the paper to increase clarity of our findings, and rewrote that paragraph in the results to read as follows: While there was a lot of variation among females in the percent of bloody pixels present per region (Fig 2), the left and/or right uterine horns had the highest percentage of bloody pixels in 6 of 10 females, while the cervix and uterus had the highest percentages in 3 of 10 females (data in S1 Table). The vagina had the lowest evidence of blood in the reproductive tract in both mated categories but one mated female (F036), had blood only in the hymen and vagina and none in the cervix, uterus or uterine horns (Fig 2, all data in S1 Table, and photos in S1 Fig 1). We also added the following sentences to the discussion: This suggests that for most of the average 20 minute copulation, the tip of the penis is likely in one or the other uterine horn rather than in the vagina. One exception to this pattern was female 36, where blood was only observed in the hymen and vagina, and none in the cervix or uterus/uterine horns. While this female mated for 30 minutes, it is possible that the penis was unable to pass through the cervix during this time, an observation that is supported by the small quantity of sperm found in her oviducts the day after mating. Attachments Attachment Submitted filename: Response to reviewers Brennan et al.docx https://doi.org/10.1371/journal.pone.0295882.r006
1 Apr 2024 Decision Letter - Jayonta Bhattacharjee, Editor Intra-horn insemination in the alpaca Vicugna pacos: Copulatory wounding and deep sperm deposition PONE-D-23-40086R3 Dear Dr. Brennan, 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, Jayonta Bhattacharjee Academic Editor PLOS ONE Additional Editor Comments (optional): Reviewers' comments: https://doi.org/10.1371/journal.pone.0295882.r007
Formally Accepted
5 Apr 2024 Acceptance Letter - Jayonta Bhattacharjee, Editor PONE-D-23-40086R3 PLOS ONE Dear Dr. Brennan, 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 If revisions are needed, the production department will contact you directly to resolve them. If no revisions are needed, you will receive an email when the publication date has been set. At this time, we do not offer pre-publication proofs to authors during production of the accepted work. Please keep in mind that we are working through a large volume of accepted articles, so please give us a few weeks 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. Jayonta Bhattacharjee Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0295882.r008

Open letter on the publication of peer review reports

PLOS recognizes the benefits of transparency in the peer review process. Therefore, we enable the publication of all of the content of peer review and author responses alongside final, published articles. Reviewers remain anonymous, unless they choose to reveal their names.

We encourage other journals to join us in this initiative. We hope that our action inspires the community, including researchers, research funders, and research institutions, to recognize the benefits of published peer review reports for all parts of the research system.

Learn more at ASAPbio .