Hidden diversity of Trypanosomatidae (Protozoa: Kinetoplastea) in bats from an urban park in Brazil

Mariana Alves Lima; Marcela Elisa Vaz; Jennifer Emanuelle Ferreira; Ana Cristina Vianna Mariano da Rocha Lima; Débora Cristina Capucci; Sônia Aparecida Talamoni; Felipe Dutra-Rêgo; José Dilermando Andrade Filho

doi:10.1371/journal.pone.0319781

Peer Review History

Original SubmissionOctober 14, 2024
14 Oct 2024 Author Response https://doi.org/10.1371/journal.pone.0319781.r001
22 Dec 2024 Decision Letter - Vinícius Belo, Editor PONE-D-24-46291Hidden diversity of Trypanosomatidae (Protozoa: Kinetoplastea) in bats from an urban park in BrazilPLOS ONE Dear Dr. Dutra-Rêgo, 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 05 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. 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Additionally, consider including information and analyses related to the ecological profile, as suggested. [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: Partly Reviewer #2: Partly Reviewer #3: Yes ******** 2. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: N/A Reviewer #2: N/A 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: No 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: The work "Hidden diversity of Trypanosomatidae (Protozoa: Kinetoplastea) in bats from an urban park in Brazil" provides information about the occurrence of trypanosomatids in bats from an urban park in Belo Horizonte, Minas Gerais, Brazil. While the data is interesting as a record of zoonotic parasites in the study site, the scope of the information generated is limited to a national or regional level. The sample size is very small, and only a few bat species were examined. In Brazil, there are various studies on trypanosomatids in bats. In fact, infection with Leishmania in the bat species from this study has already been reported in Brazil and other South American countries (see Leishmania species infection of bats: A systematic review. Acta Tropica). No additional ecological or epidemiological information about these parasites in bats is provided. I suggest submitting the work to a national or regional journal. Reviewer #2: The study under review clearly presents the main research question, which is to identify the presence of trypanosomatids in bats captured in Mangabeiras Municipal Park, Belo Horizonte, Brazil. The main findings, including the detection of different species of Leishmania and Trypanosoma, as well as the evaluation of the integrity of the extracted DNA, are well defined. The manuscript is scientifically solid, addresses a relevant topic for public health and ecology, and contributes to the understanding of the role of urban bats as potential reservoirs of parasites that cause neglected diseases, such as leishmaniasis and Chagas disease. However, there are aspects that could be improved to maximize the scientific impact and clarity of the work. Points for Improvement 1. Abstract and Introduction Contextualization and justification: Although the authors highlighted the relevance of ecological studies involving bats, it is not clear how this study advances or differs from previous works. It is recommended to: - Include a more explicit analysis of the knowledge gap this work addresses. - Add information about outbreaks or human cases in Belo Horizonte, connecting the findings to local epidemiology and comparing them with studies from other regions of Brazil or the world. References: While relevant, it would be interesting to establish a more direct connection between recent studies (last five years) and the specific focus of the research to strengthen the justification. Keywords: Including terms such as "epidemiology" or "zoonotic reservoirs" could improve the indexing and reach of the article. 2. Connection Between Methodology and Objective Methodological explanation: The introduction addresses life cycles and transmission methods of the parasites but does not link these elements to the use of Nested-PCR and other techniques employed in the study. It is recommended to explain why the chosen methodologies are suitable for detecting trypanosomatids in bats. 3. Methodological Challenges and Data Presentation Sample contamination: The contamination rate (32.2%) is acknowledged as a limitation, but it would be useful to explore more thoroughly how this contamination impacted the results. Data inconsistencies: - In the abstract, contamination is described as 31.5%, while in the results section, it is mentioned as 32.2%. This discrepancy should be corrected. - In Table 2, the detection of trypanosomatids is reported in eight animals, but the text mentions nine. This error requires careful revision. 4. Discussion on Vectors Detailing vectors: The relationship among bats, sand flies, and kissing bugs lacks sufficient detail, especially regarding potential transmission routes. It is recommended to expand this section, considering its relevance to zoonotic cycles. 5. Writing Revision Language correction: Review the manuscript's writing in English, especially in Figure 1 and lines 8-11, which contain text written in Portuguese. It is recommended to adapt these sections to English to ensure consistency and standardization throughout the document. Reviewer #3: Dear Authors, Thank you for the opportunity to review the manuscript titled "Hidden diversity of Trypanosomatidae (Protozoa: Kinetoplastea) in bats from an urban park in Brazil." The study addresses a relevant and timely topic in parasitology, providing valuable data on the diversity of trypanosomatids in bats and their potential ecological and epidemiological implications. However, I identified several areas where improvements could be made to enhance the clarity and impact of the manuscript. Below are some minor revisions: Results Line 169: “The detection of trypanosomatids through the V7V8 Nested-PCR revealed nine positive animals.” Change to "eight positive animals" or "nine positive samples." Line 170: Fig. 2A: This does not match the nine positive animals; however, changing to "nine positive samples from six animals" would be correct. The sequencing analysis is absent; including a dendrogram or phylogenetic tree to locate the sequences of the trypanosomatids found would strengthen the manuscript. Note that sequencing results are not discussed in the text. I believe these suggestions will contribute to improving the manuscript, making it more robust and clear. I congratulate the authors on their work. Sincerely. ****** 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: No 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.0319781.r002
Revision 1
20 Jan 2025 Author Response Review Comments to the Author Reviewer #1: General comment: The work "Hidden diversity of Trypanosomatidae (Protozoa: Kinetoplastea) in bats from an urban park in Brazil" provides information about the occurrence of trypanosomatids in bats from an urban park in Belo Horizonte, Minas Gerais, Brazil. While the data is interesting as a record of zoonotic parasites in the study site, the scope of the information generated is limited to a national or regional level. The sample size is very small, and only a few bat species were examined. In Brazil, there are various studies on trypanosomatids in bats. In fact, infection with Leishmania in the bat species from this study has already been reported in Brazil and other South American countries (see Leishmania species infection of bats: A systematic review. Acta Tropica). No additional ecological or epidemiological information about these parasites in bats is provided. I suggest submitting the work to a national or regional journal. Authors’ comment: Thank you for your comments. We understand that three major points were raised, and we would like to address each of them below: 1- Sample size: While we acknowledge that the sample size could be larger, we respectfully disagree with the characterization of it as “very small”. In prospective studies like ours, analyzing the highest number of samples possible is ideal to ensure representativeness. However, in urban areas such as the study site, the bat population density tends to be lower compared to wild and rural areas, which naturally impacts sample availability. Additionally, logistical challenges, including restrictions imposed by the COVID-19 pandemic during the study period, limited the feasibility of additional collections. It is important to note that sample sizes in studies on trypanosomatids in bats vary widely in the literature, with some studies reporting even smaller sample sizes than ours. We believe the data presented here provides a meaningful snapshot of the local ecological scenario, acknowledging that additional species of bats and trypanosomatids might not have been captured. This limitation, while recognized, does not diminish the value of the findings. 2- Presence of Leishmania infections already reported in Brazil and absence of ecological/epidemiological information: As shown in Table 2, our study does not focus solely on Leishmania infections, which were reported in 6 out of 9 bats. We also documented infections by Trypanosoma sp. Neobat in 3 bats. This finding is noteworthy, as there is limited information available on T. sp. Neobat, particularly in urban areas. The presence of this parasite in a protected urban area of Belo Horizonte represents a novel contribution to the field. Additionally, this is the first well-documented study on bat ecology in the protected urban areas of Belo Horizonte investigating trypanosomatid infections, further reinforcing its novelty. Regarding the ecological and epidemiological data, we have expanded this section in the revised manuscript to include more context on these aspects (lines 71-88). However, as acknowledged, knowledge gaps regarding T. sp. Neobat remain, as highlighted by several authors (e.g., Alves et al., 2021; DOI: https://doi.org/10.1016/j.ijppaw.2021.09.003). 3- Regional impact of the results: While the findings have a regional focus, we believe they provide valuable contributions to the broader understanding of trypanosomatids in bats. For instance, we expanded the geographical distribution and ecological knowledge of T. sp. Neobat, recording it for the first time in the state of Minas Gerais. This adds to the limited literature on this parasite, particularly in urban settings, and is of relevance to high-impact journals such as PLOS ONE. Reviewer #2: General comment: The study under review clearly presents the main research question, which is to identify the presence of trypanosomatids in bats captured in Mangabeiras Municipal Park, Belo Horizonte, Brazil. The main findings, including the detection of different species of Leishmania and Trypanosoma, as well as the evaluation of the integrity of the extracted DNA, are well defined. The manuscript is scientifically solid, addresses a relevant topic for public health and ecology, and contributes to the understanding of the role of urban bats as potential reservoirs of parasites that cause neglected diseases, such as leishmaniasis and Chagas disease. However, there are aspects that could be improved to maximize the scientific impact and clarity of the work. Specific comments: Abstract and Introduction Contextualization and justification: Although the authors highlighted the relevance of ecological studies involving bats, it is not clear how this study advances or differs from previous works. It is recommended to: - Include a more explicit analysis of the knowledge gap this work addresses. Authors’ comment: We included your suggestion in this new version of the manuscript (lines 71-88). - Add information about outbreaks or human cases in Belo Horizonte, connecting the findings to local epidemiology and comparing them with studies from other regions of Brazil or the world. Authors’ comment: We included your suggestion in this new version of the manuscript (lines 83-85). References: While relevant, it would be interesting to establish a more direct connection between recent studies (last five years) and the specific focus of the research to strengthen the justification. Authors’ comment: We have included your suggestion in this new version of the manuscript. Keywords: Including terms such as "epidemiology" or "zoonotic reservoirs" could improve the indexing and reach of the article. Authors’ comment: We have included "Epidemiology" and "Disease Reservoirs" as keywords. The term "zoonotic reservoir" is not a MeSH term, and therefore, we have avoided using it as a keyword. Connection Between Methodology and Objective Methodological explanation: The introduction addresses life cycles and transmission methods of the parasites but does not link these elements to the use of Nested-PCR and other techniques employed in the study. It is recommended to explain why the chosen methodologies are suitable for detecting trypanosomatids in bats. Authors’ comment: We have included your suggestion in this revised version of the manuscript (lines 63-66). Methodological Challenges and Data Presentation Sample contamination: The contamination rate (32.2%) is acknowledged as a limitation, but it would be useful to explore more thoroughly how this contamination impacted the results. Authors’ comment: We have incorporated your suggestion into the results section (lines 194-197) and the final paragraph of the discussion, where the study's limitations are addressed (lines 286-293). Data inconsistencies: - In the abstract, contamination is described as 31.5%, while in the results section, it is mentioned as 32.2%. This discrepancy should be corrected. Authors’ comment: This discrepancy has now been corrected. - In Table 2, the detection of trypanosomatids is reported in eight animals, but the text mentions nine. This error requires careful revision. Authors’ comment: Table 2 has been updated and corrected in this revised version of the manuscript. Discussion on Vectors Detailing vectors: The relationship among bats, sand flies, and kissing bugs lacks sufficient detail, especially regarding potential transmission routes. It is recommended to expand this section, considering its relevance to zoonotic cycles. Authors’ comment: In this revised version of the manuscript, we have expanded the discussion on the role of potential vectors in transmitting trypanosomatids (lines 256–277). Writing Revision Language correction: Review the manuscript's writing in English, especially in Figure 1 and lines 8-11, which contain text written in Portuguese. It is recommended to adapt these sections to English to ensure consistency and standardization throughout the document. Authors’ comment: The manuscript has been thoroughly reviewed for English language accuracy. Reviewer #3: General comment: Thank you for the opportunity to review the manuscript titled "Hidden diversity of Trypanosomatidae (Protozoa: Kinetoplastea) in bats from an urban park in Brazil." The study addresses a relevant and timely topic in parasitology, providing valuable data on the diversity of trypanosomatids in bats and their potential ecological and epidemiological implications. However, I identified several areas where improvements could be made to enhance the clarity and impact of the manuscript. Specific comments: Line 169: “The detection of trypanosomatids through the V7V8 Nested-PCR revealed nine positive animals.” Change to "eight positive animals" or "nine positive samples." Authors’ comment: This paragraph has been changed (lines 187-188). Line 170: Fig. 2A: This does not match the nine positive animals; however, changing to "nine positive samples from six animals" would be correct. Authors’ comment: Figure 2, panels A and B, represent agarose gel electrophoresis of V7V8 and gamma-actin PCR products, respectively, with a subset of positive samples displayed. The sequencing analysis is absent; including a dendrogram or phylogenetic tree to locate the sequences of the trypanosomatids found would strengthen the manuscript. Note that sequencing results are not discussed in the text. Authors’ comment: Thank you for your comment. All PCR-positive samples were 100% identical to the corresponding sequences in GenBank. In this context, constructing a phylogenetic tree would not significantly add novelty to the manuscript, as the phylogenetic positions of all parasites identified in our study are already well-established. Typically, sequencing data in such cases are presented descriptively. However, we have included Figure 3 with the phylogenetic analysis for further clarity and completeness. Attachments Attachment Submitted filename: Response to editor and reviewers.docx https://doi.org/10.1371/journal.pone.0319781.r003
6 Feb 2025 Decision Letter - Vinícius Belo, Editor PONE-D-24-46291R1Hidden diversity of Trypanosomatidae (Protozoa: Kinetoplastea) in bats from an urban park in BrazilPLOS ONE Dear Dr. Dutra-Rêgo, 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 23 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, Vinícius Silva Belo 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. [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 #2: All comments have been addressed Reviewer #3: 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 #2: Yes Reviewer #3: Yes ****** 3. Has the statistical analysis been performed appropriately and rigorously? Reviewer #2: N/A Reviewer #3: 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 #2: Yes Reviewer #3: 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 #2: No Reviewer #3: 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 #2: The manuscript "Hidden diversity of Trypanosomatidae in bats from an urban park in Brazil" presents a solid study on trypanosomatid diversity in urban bats in Belo Horizonte. The authors addressed previous concerns, enhancing the manuscript’s clarity, methodology, and scientific rigor. The study contributes valuable insights into the role of bats as reservoirs for diseases like leishmaniasis and Chagas disease. Specific comments: Abstract & Introduction: The knowledge gap is clearly defined (lines 71–88), with added local epidemiological data (lines 83–85) and relevant recent references. Keywords were updated appropriately. Methodology: The rationale for using Nested-PCR is well-explained (lines 63–66). Capture methods and ethical protocols are detailed, and the impact of the 32.2% contamination rate is thoughtfully discussed (lines 194–197, 286–293). Results & Data Consistency: Data inconsistencies have been corrected, including contamination rates and Table 2 updates. Tables and figures are clear and accurate. Discussion on Vectors: Expanded discussion on vector roles (lines 256–277), with a minor suggestion to add more references on triatomine-bat interactions. Language & Style: The language issues have been addressed, with minor suggestions to improve the text's fluency. Additionally, some sections are still written in Portuguese and should be translated to ensure consistency throughout the manuscript. Recommendation: Accept with Minor Revisions. The manuscript is well-prepared, scientifically robust, and contributes meaningful data. Minor language adjustments and additional references will further strengthen the paper. Reviewer #3: (No Response) ****** 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 #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.0319781.r004
Revision 2
6 Feb 2025 Author Response Reviewer #2: Accept with Minor Revisions. The manuscript is well-prepared, scientifically robust, and contributes meaningful data. Minor language adjustments and additional references will further strengthen the paper. Authors’ comments: We conducted a thorough review of the manuscript, addressing typographical errors and misspellings. Additionally, we have incorporated additional references related to triatomine-bat interactions, as requested. Attachments Attachment Submitted filename: Response_to_editor_and_reviewers_auresp_2.docx https://doi.org/10.1371/journal.pone.0319781.r005
9 Feb 2025 Decision Letter - Vinícius Belo, Editor Hidden diversity of Trypanosomatidae (Protozoa: Kinetoplastea) in bats from an urban park in Brazil PONE-D-24-46291R2 Dear Dr. Dutra-Rêgo, 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, Vinícius Silva Belo Academic Editor PLOS ONE Additional Editor Comments (optional): Congratulations to the authors on their insightful work. It is a pleasure to accept your manuscript for publication! Reviewers' comments: https://doi.org/10.1371/journal.pone.0319781.r006
Formally Accepted
Acceptance Letter - Vinícius Belo, Editor PONE-D-24-46291R2 PLOS ONE Dear Dr. Dutra-Rêgo, 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. Vinícius Silva Belo Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0319781.r007

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