Dear Mr. Palmer-Young,

Thank you very much for submitting your manuscript "Can floral nectars reduce transmission of Leishmania?" for consideration at PLOS Neglected Tropical Diseases. As with all papers reviewed by the journal, your manuscript was reviewed by members of the editorial board and by several independent reviewers. In light of the reviews (below this email), we would like to invite the resubmission of a significantly-revised version that takes into account the reviewers' comments.

Please, verify the comments of the reviewers carefully.

We cannot make any decision about publication until we have seen the revised manuscript and your response to the reviewers' comments. Your revised manuscript is also likely to be sent to reviewers for further evaluation.

When you are ready to resubmit, please upload the following:

[1] A letter containing a detailed list of your responses to the review comments and a description of the changes you have made in the manuscript. Please note while forming your response, if your article is accepted, you may have the opportunity to make the peer review history publicly available. The record will include editor decision letters (with reviews) and your responses to reviewer comments. If eligible, we will contact you to opt in or out.

[2] Two versions of the revised manuscript: one with either highlights or tracked changes denoting where the text has been changed; the other a clean version (uploaded as the manuscript file).

Important additional instructions are given below your reviewer comments.

Please prepare and submit your revised manuscript within 60 days. If you anticipate any delay, please let us know the expected resubmission date by replying to this email. Please note that revised manuscripts received after the 60-day due date may require evaluation and peer review similar to newly submitted manuscripts.

Thank you again for your submission. We hope that our editorial process has been constructive so far, and we welcome your feedback at any time. Please don't hesitate to contact us if you have any questions or comments.

Sincerely,

Claudia Ida Brodskyn

Associate Editor

PLOS Neglected Tropical Diseases

Alvaro Acosta-Serrano

Deputy Editor

PLOS Neglected Tropical Diseases

***********************

Please, verify the comments of the reviewers carefully.

Reviewer's Responses to Questions

Key Review Criteria Required for Acceptance?

As you describe the new analyses required for acceptance, please consider the following:

Methods

-Are the objectives of the study clearly articulated with a clear testable hypothesis stated?

-Is the study design appropriate to address the stated objectives?

-Is the population clearly described and appropriate for the hypothesis being tested?

-Is the sample size sufficient to ensure adequate power to address the hypothesis being tested?

-Were correct statistical analysis used to support conclusions?

-Are there concerns about ethical or regulatory requirements being met?

Reviewer #1: As this is a meta-analysis their is no testable hypothesis. So the other questions are irrelevant

Reviewer #2: This section methods is fine for me except some points to address. Please find below my comments

1- Line 101-102 (Ref 27), could you explain how flavonoid glycosides are less effective than aglycones?

Reviewer #3: The study design was clear, however it is not strong enough support the hypothesis raised in this study.

--------------------

Results

-Does the analysis presented match the analysis plan?

-Are the results clearly and completely presented?

-Are the figures (Tables, Images) of sufficient quality for clarity?

Reviewer #1: As this is a compilation of other scientists' research there was no analysis plan to begin with.

The results are clearly stated.

Figures:

Figure 1 A: There are no values even close to 1,000 (1e+03), so why have a graph that goes to 1e+05 (100,000)? B: Is very busy and hard to read – can it be simplified?

Figures 2 & 3. N.B. fig. 2 uses 1e + scale, while fig 3 uses 0.1 to 100. The second is so much easier to read.

Table

Table includes data on amastigotes, a stage in the life cycle of the parasite that is very transitory in the sand fly.

Reviewer #2: Good, but is the concentration of floral resources that you use the same concentration that could Phlobotomus ingest in the wild?

Reviewer #3: Concentrations evaluated here are based only in vitro exposition of the parasite to compounds, ignoring their effect during Leishmania-sand fly interactions, i.e., ignoring the behavior and physiology of the insect host, in which is essential to the potential for the vector control.

--------------------

Conclusions

-Are the conclusions supported by the data presented?

-Are the limitations of analysis clearly described?

-Do the authors discuss how these data can be helpful to advance our understanding of the topic under study?

-Is public health relevance addressed?

Reviewer #1: The conclusions are adequate, but the limitations are (i.e. lack of field data) not mentioned.

No evidence is presented that sand flies actually feed on the various plants mentioned.

In the field of Public Health the authors wrote that their work could lead to an environmentally friendly control of trypanosomatid threats to global health.

Reviewer #2: Good

Reviewer #3: The compilation done here would be extremely helpful as a screening process to test experimentally these compounds in the insect vector. Such conclusion would be more restrictive, but certainly more accurate to the data presented.

--------------------

Editorial and Data Presentation Modifications?

Use this section for editorial suggestions as well as relatively minor modifications of existing data that would enhance clarity. If the only modifications needed are minor and/or editorial, you may wish to recommend “Minor Revision” or “Accept”.

Reviewer #1: Lines 13 and 53: Insect-vectored Leishmania are the second-most debilitating of human parasites worldwide. The actual quote is “Among parasitic infections, this disease is responsible for the highest number of disability adjusted life years (a measure of health burden) after malaria.” (Lymphatic Filariasis is surely more debilitating than cutaneous leishmaniasis caused by Leishmania major). While the authors repeat several times this health burden, it is somewhat over empathized.

Line 54 The CDC numbers: For cutaneous leishmaniasis, estimates of the number of new cases per year have ranged from approximately 700,000 to 1.2 million or more. For visceral leishmaniasis, the estimated number of new cases per year may have decreased to <100,000. WHO numbers are less, so perhaps “reported” should be cautiously used.

Line 66: “floral nectar appears to be a preferred food source” Perhaps it should be noted that this study was carried out in a lush irrigated farming village surrounded by desert.

Line 137 and Discussion: Is there any evidence that sand flies feed actually on for example (Dicentra eximia, Brassica napus, Helianthus annuus, or Thymus vulgaris)? Examples of mesocosm would augment the meta-analysis. A real world example of this would be “DNA barcode for the identification of the sand fly Lutzomyia longipalpis plant feeding preferences in a tropical urban environment”: Leonardo H G de M Lima et al Sci Rep, 2016 Jul 20;6: 29742 or if the authors comb the literature they can find Suaeda asphaltica as a sand fly food source or Cameron’s work in South America on bean plants as actual sand fly food or Alexander B and Usma MC, study in Colombian coffee fields.

Could it be that the compounds the authors are promoting would make better pharmaceutical products?

While the MS is partly speculative in nature, the inclusion of some of the references add little to overall question.

Other references that may interest the authors for their discussion:

Lectins and toxins in the plant diet of Phlebotomus papatasi (Diptera: Psychodidae) can kill Leishmania major promastigotes in the sandfly and in culture. Jacobson RL, Schlein Y. Ann Trop Med Parasitol. 1999 Jun;93 (4):351-6.

Sand fly feeding on noxious plants: a potential method for the control of leishmaniasis Y. Schlein, R L Jacobson, G C Müller Am J Trop Med Hyg. 2001 Oct; 65(4):300-3

While the MS is partly speculative in nature, the inclusion of some of the references adds little to overall question?

Reviewer #2: Good

Reviewer #3: (No Response)

--------------------

Summary and General Comments

Use this section to provide overall comments, discuss strengths/weaknesses of the study, novelty, significance, general execution and scholarship. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. If requesting major revision, please articulate the new experiments that are needed.

Reviewer #1: The MS reviews the research of others on the in vitro screening of various extracts of nectar and pollen on Leishmania species and discuss the use of secondary metabolites in the control of the infections in the sand fly vector.

The inclusion of known plant food sources would improve the MS

Reviewer #2: Phlebotomus take the nectar as nutrition. The effects of components in nectar are various. This study focuses on floral nectars and Leishmania transmission. The conclusion is that planting phytochemical-rich floral resources or phytochemically enriched baits could reduce Leishmania loads in vectors, providing an environmentally friendly complement to existing means of disease control. With these findings, there is a great hope to come over some diseases as Leishmaniasis without chemical solution, using natural environmental resources.

The manuscript how it is standing now, is well written, clear and shows good results to fight against the Leishmania. However, to improve the manuscript some points raised below need to be more detailed.

1- Line 117-118 (Ref 13), in this paper they have shown that some plant species reduce the transmission of malaria and other plants maintain the transmission, so, could you consider this in the discussion?

2- In your references 2, 12, 17, 23 there is no DOI as well as in all your references in Supporting information 1, please add these details

3- Some species scientific names in your references are not italicized, you should correct that too

Reviewer #3: The information discussed in this study is certainly interesting, but it is not being consistent with the broader potential speculated by authors. I suggest authors either support the study with some experimental tests of nectar compounds on insects or restrict claims to avoid the speculative character.

--------------------

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Reviewer #1: No

Reviewer #2: No

Reviewer #3: No

Figure 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. 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 us at figures@plos.org.

Data Requirements:

Please note that, as a condition of publication, PLOS' data policy requires that you make available all data used to draw the conclusions outlined in your manuscript. Data must be deposited in an appropriate repository, included within the body of the manuscript, or uploaded as supporting information. This includes all numerical values that were used to generate graphs, histograms etc.. For an example see here: http://www.plosbiology.org/article/info%3Adoi%2F10.1371%2Fjournal.pbio.1001908#s5.

Reproducibility:

To enhance the reproducibility of your results, we recommend that you deposit your laboratory protocols in protocols.io, where a protocol can be assigned its own identifier (DOI) such that it can be cited independently in the future. Additionally, PLOS ONE offers an option to publish peer-reviewed clinical study protocols. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols

Dear Palmer-Young,

Thank you very much for submitting your manuscript "Can floral nectars reduce transmission of Leishmania?" for consideration at PLOS Neglected Tropical Diseases. As with all papers reviewed by the journal, your manuscript was reviewed by members of the editorial board and by several independent reviewers. In light of the reviews (below this email), we would like to invite the resubmission of a significantly-revised version that takes into account the reviewers' comments.

Although the modifications in the text have improved the manuscript, there are still some alterations that deserve attention by the authors.

We cannot make any decision about publication until we have seen the revised manuscript and your response to the reviewers' comments. Your revised manuscript is also likely to be sent to reviewers for further evaluation.

When you are ready to resubmit, please upload the following:

[1] A letter containing a detailed list of your responses to the review comments and a description of the changes you have made in the manuscript. Please note while forming your response, if your article is accepted, you may have the opportunity to make the peer review history publicly available. The record will include editor decision letters (with reviews) and your responses to reviewer comments. If eligible, we will contact you to opt in or out.

[2] Two versions of the revised manuscript: one with either highlights or tracked changes denoting where the text has been changed; the other a clean version (uploaded as the manuscript file).

Important additional instructions are given below your reviewer comments.

Please prepare and submit your revised manuscript within 60 days. If you anticipate any delay, please let us know the expected resubmission date by replying to this email. Please note that revised manuscripts received after the 60-day due date may require evaluation and peer review similar to newly submitted manuscripts.

Thank you again for your submission. We hope that our editorial process has been constructive so far, and we welcome your feedback at any time. Please don't hesitate to contact us if you have any questions or comments.

Sincerely,

Claudia Ida Brodskyn

Associate Editor

PLOS Neglected Tropical Diseases

Alvaro Acosta-Serrano

Deputy Editor

PLOS Neglected Tropical Diseases

***********************

Although the modifications inthe text have improved the manuscript, there are still some alterations that deserve attention by the authors.

Reviewer's Responses to Questions

Key Review Criteria Required for Acceptance?

As you describe the new analyses required for acceptance, please consider the following:

Methods

-Are the objectives of the study clearly articulated with a clear testable hypothesis stated?

-Is the study design appropriate to address the stated objectives?

-Is the population clearly described and appropriate for the hypothesis being tested?

-Is the sample size sufficient to ensure adequate power to address the hypothesis being tested?

-Were correct statistical analysis used to support conclusions?

-Are there concerns about ethical or regulatory requirements being met?

Reviewer #1: (No Response)

Reviewer #2: Good, very clear to me now

Reviewer #3: As previously mentioned the study is clear and presents a satisfactory design.

--------------------

Results

-Does the analysis presented match the analysis plan?

-Are the results clearly and completely presented?

-Are the figures (Tables, Images) of sufficient quality for clarity?

Reviewer #1: (No Response)

Reviewer #2: Good

Reviewer #3: Data presented previously was clear and modifications have not big effect in the current manuscript.

--------------------

Conclusions

-Are the conclusions supported by the data presented?

-Are the limitations of analysis clearly described?

-Do the authors discuss how these data can be helpful to advance our understanding of the topic under study?

-Is public health relevance addressed?

Reviewer #1: (No Response)

Reviewer #2: Good

Reviewer #3: In my view, the conclusion of this work still remain very broad considering the data based only in vitro tests to discuss a bigger question, a more restrictive statement would be more plausible.

--------------------

Editorial and Data Presentation Modifications?

Use this section for editorial suggestions as well as relatively minor modifications of existing data that would enhance clarity. If the only modifications needed are minor and/or editorial, you may wish to recommend “Minor Revision” or “Accept”.

Reviewer #1: (No Response)

Reviewer #2: Good

Reviewer #3: I acknowledge all changes performed by the authors in the text, it improved considerably the manuscript, however I still would recommend some "major revision" in this work.

--------------------

Summary and General Comments

Use this section to provide overall comments, discuss strengths/weaknesses of the study, novelty, significance, general execution and scholarship. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. If requesting major revision, please articulate the new experiments that are needed.

Reviewer #1: The authors have used the remarks of the referees to refine their MS. It has been improved.

Reviewer #2: Good from my point of view

Reviewer #3: Despite all modifications performed, the hypothesis raised in this work still consider a large knowledge in nectar compounds and their effect in the parasite in vitro, it has poor information related to the vector interaction, in which is essential considering to the potential claimed in this manuscript.

I still recommend authors discuss more about the physiology and behaviour of the insect that could justify the hypothesis. Even simple details regarding the insect could make all the difference and should be not selectively ignored. For instance, if the nectar amount taken by the insect would enough to affect the parasite considering the in vitro results presented here?

Please check here some papers that might help :

Ferreira, T.N., Pita-Pereira, D., Costa, S.G. et al. Transmission blocking sugar baits for the control of Leishmania development inside sand flies using environmentally friendly beta-glycosides and their aglycones. Parasites Vectors 11, 614 (2018). https://doi.org/10.1186/s13071-018-3122-z

Ferreira ME, de Arias AR, Yaluff G, de Bilbao NV, Nakayama H, Torres S, et al. Antileishmanial activity of furoquinolines and coumarins from Helietta apiculata. Phytomedicine. 2010;17:375–8.

Gontijo NF, Melo MN, Riani EB, Aleida-Silva S, Mares-Guia ML. Glycosidases in Leishmania and their importance for Leishmania in phlebotomine sand flies with special reference to purification and characterization of a sucrase. Exp Parasitol. 1996;83:117–24.

Jacobson RL, Schlein Y, Eisenberger CL. The biological function of sand fly and Leishmania glycosidases. Med Microbiol Immunol. 2001;190:51–5

Dillon RJ, EEl K. Carbohydrate digestion in sand flies: α-glucosidase activity in the midgut of Phlebotomus langeroni. Comp Biochem Physiol. 1997;116B:35–40.

--------------------

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

Figure 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. 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 us at figures@plos.org.

Data Requirements:

Please note that, as a condition of publication, PLOS' data policy requires that you make available all data used to draw the conclusions outlined in your manuscript. Data must be deposited in an appropriate repository, included within the body of the manuscript, or uploaded as supporting information. This includes all numerical values that were used to generate graphs, histograms etc.. For an example see here: http://www.plosbiology.org/article/info%3Adoi%2F10.1371%2Fjournal.pbio.1001908#s5.

Reproducibility:

To enhance the reproducibility of your results, we recommend that you deposit your laboratory protocols in protocols.io, where a protocol can be assigned its own identifier (DOI) such that it can be cited independently in the future. Additionally, PLOS ONE offers an option to publish peer-reviewed clinical study protocols. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols

Dear Palmer-Young,

Thank you very much for submitting your manuscript "Can floral nectars reduce transmission of Leishmania?" for consideration at PLOS Neglected Tropical Diseases. As with all papers reviewed by the journal, your manuscript was reviewed by members of the editorial board and by several independent reviewers. The reviewers appreciated the attention to an important topic. Based on the reviews, we are likely to accept this manuscript for publication, providing that you modify the manuscript according to the review recommendations.

Dear authors,

The manuscript is improved a lot, but two reviewers pointed out some minor issues that deserve your attention.

Please, answer these points and send the manuscript back to be analyzed again.

Please prepare and submit your revised manuscript within 30 days. If you anticipate any delay, please let us know the expected resubmission date by replying to this email.

When you are ready to resubmit, please upload the following:

[1] A letter containing a detailed list of your responses to all review comments, and a description of the changes you have made in the manuscript.

Please note while forming your response, if your article is accepted, you may have the opportunity to make the peer review history publicly available. The record will include editor decision letters (with reviews) and your responses to reviewer comments. If eligible, we will contact you to opt in or out

[2] Two versions of the revised manuscript: one with either highlights or tracked changes denoting where the text has been changed; the other a clean version (uploaded as the manuscript file).

Important additional instructions are given below your reviewer comments.

Thank you again for your submission to our journal. We hope that our editorial process has been constructive so far, and we welcome your feedback at any time. Please don't hesitate to contact us if you have any questions or comments.

Sincerely,

Claudia Ida Brodskyn

Associate Editor

PLOS Neglected Tropical Diseases

Alvaro Acosta-Serrano

Deputy Editor

PLOS Neglected Tropical Diseases

***********************

Dear all,

The manuscript is improved a lot, but two reviewers pointed out some issues that deserve your attention.

Please, answer these points and send the manuscript back to be analyzed agains.

Reviewer's Responses to Questions

Key Review Criteria Required for Acceptance?

As you describe the new analyses required for acceptance, please consider the following:

Methods

-Are the objectives of the study clearly articulated with a clear testable hypothesis stated?

-Is the study design appropriate to address the stated objectives?

-Is the population clearly described and appropriate for the hypothesis being tested?

-Is the sample size sufficient to ensure adequate power to address the hypothesis being tested?

-Were correct statistical analysis used to support conclusions?

-Are there concerns about ethical or regulatory requirements being met?

Reviewer #2: Good for me

Reviewer #3: Methods are clear and well described by the authors

Reviewer #4: (No Response)

--------------------

Results

-Does the analysis presented match the analysis plan?

-Are the results clearly and completely presented?

-Are the figures (Tables, Images) of sufficient quality for clarity?

Reviewer #2: Good

Reviewer #3: Result analysis is consistent with the study design and clearly explained.

Reviewer #4: see below

--------------------

Conclusions

-Are the conclusions supported by the data presented?

-Are the limitations of analysis clearly described?

-Do the authors discuss how these data can be helpful to advance our understanding of the topic under study?

-Is public health relevance addressed?

Reviewer #2: Good

Reviewer #3: Lines: 289-292

I would like just to point that some technical interspecies issues may need to be addressed here when extrapolating these results. It is unlikely that the phytochemistry from these floral compounds would be specific to sand flies. The ecological concerns would be similar to using synthetic compounds.

Reviewer #4: see below

--------------------

Editorial and Data Presentation Modifications?

Use this section for editorial suggestions as well as relatively minor modifications of existing data that would enhance clarity. If the only modifications needed are minor and/or editorial, you may wish to recommend “Minor Revision” or “Accept”.

Reviewer #2: Good

Reviewer #3: Accept

Reviewer #4: see below

--------------------

Summary and General Comments

Use this section to provide overall comments, discuss strengths/weaknesses of the study, novelty, significance, general execution and scholarship. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. If requesting major revision, please articulate the new experiments that are needed.

Reviewer #2: Good for me now, but they need to consider changing Phlebotomus langeroni in italic in reference 23 and the figure 1 in supporting information section is blurred

Reviewer #3: I acknowledge all modifications performed in the manuscript. The text is more consistent with the data analysed.

Reviewer #4: Revised version of the manuscript is significantly improved as helpful comments of reviewers were accepted. In my opinion, just few relatively minor changes are required:

Authors are aware that the sugar meal is directed first to crop (see lines 237-240). However, I am missing important information that only small droplets of sugar meal are released from the crop to the anterior (thoracic) midgut where the sugar meal is relatively quickly digested by glycosidases. This mechanism of sugar digestion may significantly decrease the effect of anti-leishmania compounds in the sugar meal. It is necessary to mention this fact either in Discussion or Conclusions.

The above mentioned fact should be also reflected by more cautious deductions in the text, for example „would be“ should be replaced by „might be“ on line 267.

Line 274: the proportion of microbiota in sand fly hindgut is negligible and unimportant in comparison to microbiota present in abdominal midgut of sand flies. Therefore, „hindgut microbiota“ should be replaced by „microbiota in abdominal midgut“.

--------------------

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

Reviewer #4: No

Figure 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. 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 us at figures@plos.org.

Data Requirements:

Please note that, as a condition of publication, PLOS' data policy requires that you make available all data used to draw the conclusions outlined in your manuscript. Data must be deposited in an appropriate repository, included within the body of the manuscript, or uploaded as supporting information. This includes all numerical values that were used to generate graphs, histograms etc.. For an example see here: http://www.plosbiology.org/article/info%3Adoi%2F10.1371%2Fjournal.pbio.1001908#s5.

Reproducibility:

To enhance the reproducibility of your results, we recommend that you deposit your laboratory protocols in protocols.io, where a protocol can be assigned its own identifier (DOI) such that it can be cited independently in the future. Additionally, PLOS ONE offers an option to publish peer-reviewed clinical study protocols. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols

References

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.

Dear Palmer-Young,

We are pleased to inform you that your manuscript 'Can floral nectars reduce transmission of Leishmania?' has been provisionally accepted for publication in PLOS Neglected Tropical Diseases.

Before your manuscript can be formally accepted you will need to complete some formatting changes, which you will receive in a follow up email. A member of our team will be in touch with a set of requests.

Please note that your manuscript will not be scheduled for publication until you have made the required changes, so a swift response is appreciated.

IMPORTANT: The editorial review process is now complete. PLOS will only permit corrections to spelling, formatting or significant scientific errors from this point onwards. Requests for major changes, or any which affect the scientific understanding of your work, will cause delays to the publication date of your manuscript.

Should you, your institution's press office or the journal office choose to press release your paper, you will automatically be opted out of early publication. We ask that you notify us now if you or your institution is planning to press release the article. All press must be co-ordinated with PLOS.

Thank you again for supporting Open Access publishing; we are looking forward to publishing your work in PLOS Neglected Tropical Diseases.

Best regards,

Claudia Ida Brodskyn

Associate Editor

PLOS Neglected Tropical Diseases

Alvaro Acosta-Serrano

Deputy Editor

PLOS Neglected Tropical Diseases

***********************************************************

After three rounds of revision, the manuscript can be accepted for publication.\\Best regards

Cláudia Brodskyn