Impact of feeding strategies on the welfare and behaviour of horses in groups: An experimental study

Marie Roig-Pons; Iris Bachmann; Sabrina Briefer Freymond

doi:10.1371/journal.pone.0325928

Peer Review History

Original SubmissionJanuary 17, 2025
17 Jan 2025 Author Response https://doi.org/10.1371/journal.pone.0325928.r001
Decision Letter - Julio Cesar de Souza, Editor PONE-D-25-03027Impact of feeding strategies on the welfare and behaviour of horses in groups: an experimental studyPLOS ONE Dear Dr. Roig-Pons, 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. ============================== Dear Sir, Can you make the reviewer's suggestions to finish the process? Best regards, ============================== Please submit your revised manuscript by Apr 04 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. 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, Julio Cesar de Souza, Ph.D. 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. When completing the data availability statement of the submission form, you indicated that you will make your data available on acceptance. We strongly recommend all authors decide on a data sharing plan before acceptance, as the process can be lengthy and hold up publication timelines. Please note that, though access restrictions are acceptable now, your entire data will need to be made freely accessible if your manuscript is accepted for publication. This policy applies to all data except where public deposition would breach compliance with the protocol approved by your research ethics board. If you are unable to adhere to our open data policy, please kindly revise your statement to explain your reasoning and we will seek the editor's input on an exemption. Please be assured that, once you have provided your new statement, the assessment of your exemption will not hold up the peer review process. 3. Your ethics statement should only appear in the Methods section of your manuscript. If your ethics statement is written in any section besides the Methods, please delete it from any other section. 4. We note that Figures 1 and 2 in your submission contain copyrighted images. All PLOS content is published under the Creative Commons Attribution License (CC BY 4.0), which means that the manuscript, images, and Supporting Information files will be freely available online, and any third party is permitted to access, download, copy, distribute, and use these materials in any way, even commercially, with proper attribution. For more information, see our copyright guidelines: http://journals.plos.org/plosone/s/licenses-and-copyright. We require you to either (1) present written permission from the copyright holder to publish these figures specifically under the CC BY 4.0 license, or (2) remove the figures from your submission: 1. You may seek permission from the original copyright holder of Figures 1 and 2 to publish the content specifically under the CC BY 4.0 license. We recommend that you contact the original copyright holder with the Content Permission Form (http://journals.plos.org/plosone/s/file?id=7c09/content-permission-form.pdf) and the following text: “I request permission for the open-access journal PLOS ONE to publish XXX under the Creative Commons Attribution License (CCAL) CC BY 4.0 (http://creativecommons.org/licenses/by/4.0/). Please be aware that this license allows unrestricted use and distribution, even commercially, by third parties. Please reply and provide explicit written permission to publish XXX under a CC BY license and complete the attached form.” Please upload the completed Content Permission Form or other proof of granted permissions as an "Other" file with your submission. In the figure caption of the copyrighted figure, please include the following text: “Reprinted from [ref] under a CC BY license, with permission from [name of publisher], original copyright [original copyright year].” 2. If you are unable to obtain permission from the original copyright holder to publish these figures under the CC BY 4.0 license or if the copyright holder’s requirements are incompatible with the CC BY 4.0 license, please either i) remove the figure or ii) supply a replacement figure that complies with the CC BY 4.0 license. Please check copyright information on all replacement figures and update the figure caption with source information. If applicable, please specify in the figure caption text when a figure is similar but not identical to the original image and is therefore for illustrative purposes only. 5. 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. [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 Reviewer #3: Yes ******** 2. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: Yes Reviewer #2: Yes Reviewer #3: 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 Reviewer #3: 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 Reviewer #3: 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: This study looked at 3 feeding stragies for horses using group housed mares, assessing impacts on behaviour and injury. The manuscript is well written with excellent detail especially in the methods and declaration of confounders due to a severe weather event. In consideration of general welfare and the impact of seasonality, please more clearly state the seasons and time periods when the study was conducted for the replicates. Especially for mares, transition to long day light under natural lighting conditions will induce cycling which could potentially alter interactions. This could be briefly touched on in the discussion or limitations. Was body weight and body condition score monitored? Could this be reported for the groups given the differences in the feed regimes this would be of considerable interest to horse owners and welfarists. Alternatively, is there an indication of dry matter intake for the group/ individual horse and a significant change between feed regimes on this? In the limitations this wasn't discussed but it would be an important inclusion for future work if not available in this study. Line 514 minor syntax error "due to the increased number of food delivery" Change to "Due to the increased number of food deliveries" Reviewer #2: I really liked the article, found a interesting study and method of observations. I have some questions: How were the injuries evaluated? The horses where taken to another place or restrained? About the disruption incident did it had any impact on the data analysis? Do the authors think it might influenced in the animals feed or behaviour? Does the authors plan to do another research where they can measure the levels of cortisol in each treatment, to link it to the animals behavour? Reviewer #3: The manuscript entitled ‘Impact of feeding strategies on the welfare and behaviour of horses in groups: an experimental study’ set out to investigate possible feeding strategies to meet the needs of horses in particular with respect to the welfare to be maintained. It is believed that the manuscript clearly presented the results obtained. The text is well presented, there are some sections that can be improved, in general it is requested to make a slight revision of the English, to shorten some sentences considered too long and to add some tables to make the text more fluent. In the introduction, the text is well written and detailed, and offers a clear explanation of the topic, there might be some details to revise especially with respect to the contextualisation of the information. In the line 45, it would be useful to elaborate and give some more references because, despite the reduced time, social interactions are important for the horse's welfare. With regard to welfare, it might be necessary to make a brief reference to what activities horses are most involved in (e.g. training, transport and other activities that undermine their welfare). Two articles are attached in this regard: Rizzo M. et al., Cortisol levels and leukocyte population values in transported and exercised horses after acupuncture needle stimulation Journal of Veterinary Behavior Volume 18, Pages 561 March 2017. Piccione G. et al., Serum lipid modification related to exercise and polyunsaturated fatty acid supplementation in jumpers and thoroughbred horses Journal of Equine Veterinary ScienceVolume 34, Issue 10, Pages 1181 - 11871 October 2014. With respect to group activities, he would emphasise and elaborate on the availability of resources. It should be made clear that not all horses are susceptible to obesity, but that there is a predisposition. In the materials and methods section, it is asked to be more specific with respect to the physical condition of the animals. Social interactions were well evaluated, perhaps a table could be presented to summarise them, in addition to the one already presented. The results were well presented, the part referring to tables and graphs contributes well to presenting them. In the discussion section, it would be necessary to make a more specific and extensive comparison with previous studies, in order to expand the bibliography already present within the study, it would be advisable to go into more detail on competitive interactions. While the results are presented and properly discussed, it would be useful to extend the discussion on how these results may influence daily animal management practices on the farm. ****** 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: Rachel Allavena Reviewer #2: No Reviewer #3: 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. https://doi.org/10.1371/journal.pone.0325928.r002
Revision 1
30 Apr 2025 Author Response B. Response to the Reviewers 1. Is the manuscript technically sound, and do the data support the conclusions? Reviewer #1: Yes Reviewer #2: Yes Reviewer #3: Yes R: We thank the three reviewers for this positive evaluation of our manuscript. ________________________________________ 2. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: Yes Reviewer #2: Yes Reviewer #3: Yes R: We thank the three reviewers for the positive evaluation of our statistical analysis ________________________________________ 3. Have the authors made all data underlying the findings in their manuscript fully available? Reviewer #1: Yes Reviewer #2: Yes Reviewer #3: Yes R: We thank the three reviewers for this positive assessment. The datasets generated and used for the analysis have been made available on Zenodo at this DOI : 10.5281/zenodo.15090821. Access is restricted at the time of review, but the data will be made public if the manuscript is accepted for publication. ________________________________________ 4. Is the manuscript presented in an intelligible fashion and written in standard English? Reviewer #1: Yes Reviewer #2: Yes Reviewer #3: Yes R: We would like to thank the three reviewers for their positive assessment of our work. ________________________________________ 5. Review Comments to the Author R: Please note that in the following answers the line numbering corresponds to that of the manuscript with the track of changes. ________________________________________ Reviewer #1 This study looked at 3 feeding stragies for horses using group housed mares, assessing impacts on behaviour and injury. The manuscript is well written with excellent detail especially in the methods and declaration of confounders due to a severe weather event. In consideration of general welfare and the impact of seasonality, please more clearly state the seasons and time periods when the study was conducted for the replicates. Especially for mares, transition to long day light under natural lighting conditions will induce cycling which could potentially alter interactions. This could be briefly touched on in the discussion or limitations. R: We deeply thank the reviewer for the very nice feedback on our manuscript. Regarding the time periods when the study was conducted, it was already stated in the first paragraph of the Material and Method “The study was conducted between February and July 2023” (L111-112). However, we took good note of the reviewer’s comment and added “Finally, data collection took place from 1 to 15 March (Repeat 1), 4 to 17 June (Repeat 2) and 3 to 16 July (Repeat 3)” (L240-241) after the description of the unforeseen event, to enhance clarity regarding the time periods. The following sentences were included in the Limitation paragraph to address this additional limitation with transparency: “Finally, this event postponed the time period of Repeat 2 and Repeat 3, leading to the first data collection in early Spring while the two remaining ones were conducted in Summer. This may have influenced the behaviour of the mares, possibly due to the presence of insects and the warm weather [77,78] or the cycling that is known to be impacted by the season [79].” (L643-646) Was body weight and body condition score monitored? Could this be reported for the groups given the differences in the feed regimes this would be of considerable interest to horse owners and welfarists. Alternatively, is there an indication of dry matter intake for the group/ individual horse and a significant change between feed regimes on this? In the limitations this wasn't discussed but it would be an important inclusion for future work if not available in this study. R: We thank the reviewer for his insightful comment. Regarding body condition, it was assessed at the beginning of the experiment both by BCS and by weighting the mares. However, we decided to rely on body weight changes to optimise objectivity by weighing the mares at the end of each Repeat. Unfortunately, our scale had a 20kg margin of error and we very rarely had a weight difference > 20kg due to the very short time between two measurements. In fact, apart from the assessment after the 2nd replicate - due to the unforeseen event and the change in feeding regime independent of the study - we had 4 and 3 horses that showed a weight difference> 20kg after the 1st and 3rd replicate respectively. Therefore, the measurements were not really meaningful (due to both the margin of error and the unforeseen event) and we decided to withdraw this part of the manuscript. To improve clarity on this issue, we have added a reference to the mares' body condition at the start of the experiment (L122-124) and a note on the problem we encountered in monitoring body weight changes (L167-171) in the Material and Methods section Regarding the dry matted intake, this was unfortunately not feasible in our experiment, as stated in our Limitations paragraph (L655-656). In the same paragraph, we have already reported the findings of Seabra et al. and DeBoer et al. (two studies with similar treatments and protocols) regarding body weight and hay ingestion.: “Seabra et al. [29] found slow-feeding and portioned feeding resulted in similar hay ingestion (1.9% of BW), whereas free-choice feeding exceeded 3% BW. DeBoer et al. [74] found a similar result (2.6% of BW with nets vs. 3.2% without) and also noted differences in BW and body condition score (BCS) with and without nets, highlighting the impact of feeding strategy on horse health metrics.” (658-660) Line 514 minor syntax error "due to the increased number of food delivery" Change to "Due to the increased number of food deliveries" R: We have corrected accordingly (L527) ________________________________________ Reviewer #2 I really liked the article, found a interesting study and method of observations. I have some questions: How were the injuries evaluated? The horses where taken to another place or restrained? About the disruption incident did it had any impact on the data analysis? Do the authors think it might influenced in the animals feed or behaviour? Does the authors plan to do another research where they can measure the levels of cortisol in each treatment, to link it to the animals behavour? R: We thank the reviewer for the positive feedback on our study and manuscript. Injuries were assessed using an adaptation of the protocol described in Zollinger et al. The mares were not removed from their group, but an experimenter was lightly restrained to the mare being assessed. We have added this information to the injuries section for clarity: “This was done by one experimenter restraining a mare while a second experimenter assessed the horse. The mares were assessed in their group to avoid any stress to the horses.” (L211-213) Regarding the effect of the disruption event, its potential impact has already been discussed in our limitation (L636-641). However, we have added a sentence to better discuss the potential impact on the mares' behaviour (feeding, aggressive behaviour and cycling). (L643-646). As for the data analysis, it was adjusted for the disruption by treating the “Repeat” variable as a fixed effect when we saw that our random components had a high variance. This is described in the Material and Methods section (L315-319). Finally, such research is not yet planned, as this project was the final experiment of the first author's PhD. However, we hope that further research will be carried out on this topic, as we strongly believe that such a study with physiological assessment would be very valuable. ________________________________________ Reviewer #3 The manuscript entitled ‘Impact of feeding strategies on the welfare and behaviour of horses in groups: an experimental study’ set out to investigate possible feeding strategies to meet the needs of horses in particular with respect to the welfare to be maintained. It is believed that the manuscript clearly presented the results obtained. The text is well presented, there are some sections that can be improved, in general it is requested to make a slight revision of the English, to shorten some sentences considered too long and to add some tables to make the text more fluent. R: We thank the reviewer for this positive comment. According to the comment, we have carefully revised the latest version of the manuscript. In addition, we would like to inform the reviewer that our previous version had been proofread by two native speakers. However, we would be willing to revise it slightly and improve the sections, if we could be told which sections is the reviewer referring to. In the introduction, the text is well written and detailed, and offers a clear explanation of the topic, there might be some details to revise especially with respect to the contextualisation of the information. R: We thank the reviewer for his comment. We have shortened a sentence that appeared too long (L84-85). As mentioned above, we would be happy to check these details if the reviewer could tell us which details could be improved. In the line 45, it would be useful to elaborate and give some more references because, despite the reduced time, social interactions are important for the horse's welfare. Regarding welfare, it might be necessary to make a brief reference to what activities horses are most involved in (e.g. training, transport and other activities that undermine their welfare). Two articles are attached in this regard: Rizzo M. et al., Cortisol levels and leukocyte population values in transported and exercised horses after acupuncture needle stimulation Journal of Veterinary Behavior Volume 18, Pages 561 March 2017. Piccione G. et al., Serum lipid modification related to exercise and polyunsaturated fatty acid supplementation in jumpers and thoroughbred horses Journal of Equine Veterinary ScienceVolume 34, Issue 10, Pages 1181 - 11871 October 2014. R We thank the reviewer for this comment. We think that, like the reviewer, social interactions are very important, despite their low frequency. However, we did not feel it was necessary to add any text on this subject, as we have already addressed it in line (L46-49) “Social interactions represent only a small proportion of a horse’s activity time-budget (3–4%), yet they are essential for the stability and cohesion of the equine social unit [6,7]. The presence of conspecifics provides not only opportunities for social interaction but also a sense of security, synchronization of rhythms, social learning for young horses, and protection from predators”. Given that the subject of this manuscript concerns the housing of equines, we do not consider it necessary to address the topic of the transport and use of equines. In addition, our experimental mares were not ridden nor transported. However, we would be willing to revise the manuscript accordingly if the reviewer provides us with a little more information on why he/she thinks it is necessary to address the topic. With respect to group activities, he would emphasise and elaborate on the availability of resources. It should be made clear that not all horses are susceptible to obesity, but that there is a predisposition. R: We agree with the reviewer that not all horses are subject to obesity, and we have indicated this in our manuscript “many horses are “easy keepers” with low energy expenditure or metabolic predispositions (…) “(L77-78). However, we do not fully understand the reviewer's request in relation to this topic. We would be willing to revise the manuscript accordingly if the reviewer clarifies his comment. In the materials and methods section, it is asked to be more specific with respect to the physical condition of the animals. R: We thank the reviewer for his perceptive comment. We have included a reference to the initial body condition score at the start of the study “In terms of body condition, at the start of the experiment all the mares had a BCS between 4 and 7 (i.e. they were all in perfect to overconditioned according the Henneke scale [40]).” (L122-124). Unfortunately, the BCS was only recorded at the beginning of the study as we relied on a scale to assess the effect of our feeding regimes on the physical condition of the horses. The mares were weighed at the start of the experiment and at the end of each treatment. However, unfortunately our scale had a 20kg margin of error and we very rarely had a weight difference > 20kg due to the very short time between two measurements. In fact, apart from the assessment after the 2nd replicate - due to the unforeseen event and the change in feeding regime independent of the study - we had 4 and 3 weight differences > 20kg after the 1st and 3rd replicate respectively. Therefore, the measurements were not meaningful (due to both the margin of error and the unforeseen event) and we decided to withdraw this part of the manuscript. To improve clarity on this issue, we have a note on the problem we encountered in monitoring body weight changes (L167-171) in the Material and Methods section. Social interactions were well evaluated, perhaps a table could be presented to summarise them, in addition to the one already presented. R: We thank the reviewer for this comment. A table summarising the different behaviours was already included in the manuscript (Table 1). However, following the reviewer's suggestion, we have now provided a detailed ethogram in a supplementary file (S1). The results were well presented, the part referring to tables and graphs contributes well to presenting them. R: We thank the reviewer for the positive feedback on our Results sections. In the discussion section, it would be necessary to make a more specific and extensive comparison with previous studies, in order to expand the bibliography already present within the study, it would be advisable to go into more detail on competitive interactions. While the results are presented and properly discussed, it would be useful to extend the discussion on how these results may influence daily animal management practices on the farm. R: We thank the reviewer for his insightful comments. We discussed our results in the light of the results of other studies, particularly those by Seabra and De Boer, which have similar protocols, to emphasise or discuss our findings. We have also added a reference to another study on agonistic interaction with hay nets “On the other hand, nets, and especially small hole hay nets may increase threats and aggression [29, 38]. This may be due to frustration, or it may indicate that hay is seen as a limited resource, thus increasing competitive interaction” (L544-547). Finally, regarding daily animal management practices on farm, as highlighted several times in the manuscript, further research is required before actual recommendation can be made, and especially research on the long-term effect of such feeding management. However, we emphasized this in a new sentence “Future research is crucial to understand the long-term effects of different feeding strategies on horse welfare, so that practical management recommendations can be accurately made and the welfare of horses on farms can be ensured.” (L562-563). ________________________________________ Attachments Attachment Submitted filename: Rebuttal Letter.docx https://doi.org/10.1371/journal.pone.0325928.r003
Decision Letter - Julio Cesar de Souza, Editor Impact of feeding strategies on the welfare and behaviour of horses in groups: an experimental study PONE-D-25-03027R1 Dear Dr. Roig-Pons, 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, Julio Cesar de Souza, Ph.D. Academic Editor PLOS ONE Additional Editor Comments (optional): Reviewers' comments: https://doi.org/10.1371/journal.pone.0325928.r004
Formally Accepted
Acceptance Letter - Julio Cesar de Souza, Editor PONE-D-25-03027R1 PLOS ONE Dear Dr. Roig-Pons, 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. Julio Cesar de Souza Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0325928.r005

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 .