Dear Dr. Coffeng,

Thank you very much for submitting your manuscript "A general framework to support cost-efficient fecal egg count method and study design choices for large-scale STH deworming programs – monitoring of therapeutic drug efficacy as a case study" 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.

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,

David Joseph Diemert, M.D.

Academic Editor

PLOS Neglected Tropical Diseases

Cinzia Cantacessi

Section Editor

PLOS Neglected Tropical Diseases

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

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: In the paragraph starting on line 184, the authors state that the stool samples were first grouped in batches of ten samples and then all samples were homogenized by stirring. It was unclear to me if the 10 samples from different individuals were mixed all together. When reading supplementary information 1, it became more clear to me that the samples were not mixed, but processed as individual samples in batches of 10. For clarity, it would be better if that was made clear in this paragraph of the main manuscript. Also, that subsamples from each of the individual samples were then processed using the three FEC methods.

Reviewer #2: The background and aims of the study, i.e. to provide an novel framework for cost-efficient study design to examine soil-transmitted-helminth monitoring and evaluation of deworming program efficacies. In my view the objectives were addressed appropriately and by considering key parameters including different diagnostic methods, parasite species, field-relevant different epidemiological scenarios and survey designs. The study population is clearly presented and appropriate to test the hypothesis that by comparing the above listed factors evidence-based recommendations can be developed for cost-efficient studies monitoring the efficacy of deworming programs. I don't feel competent to comment on the used statistical methods. I can't see any ethical or regulatory concerns.

Reviewer #3: Yes

Reviewer #4: Comments attached

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

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: The paragraph in lines 403 to 419 is confusing. Maybe some of the examples can be better displayed in table form?

Line 445, the word "redrawing" sound like you drew the graphs instead of plotting the ascaris and trichuris data.

Reviewer #2: The study results are clearly and comprehensively presented, the figures and tables are of appropriate quality, correct and informative. The presented data match the analysis plan.

Reviewer #3: Yes

Reviewer #4: Comments attached

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

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 comments.

Reviewer #2: The interpretation of the data is clear and comprehensible. The limitations (and strengths) of the study are discussed in detail in a specific paragraph of the manuscript. The whole concept of the study is to optimize the planning and conduct of deworming surveys, which also demonstrates its public health relevance.

Reviewer #3: Yes

Reviewer #4: Comments attached

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

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: In the title, the word method is missing an "s".

It´s not clear throughout the manuscript what is the exact range of subjects enrolled that were considered. In the abstract its states 10-2,000 (line 57), but in the method section (line 292) it says for a survey design a range of 100 - 5,000 was used.

Line 109 of the introduction, the word "those" is incorrectly used in this sentence.

Reviewer #2: Minor issues:

Figure 1 with respect to the Mini-FLOTAC flow chart in my view it should not read Accumulation but Flotation.

Concerning the calculation of the required time to analyse a stool sample with one of the three methods I wonder why for the FECPAG method time for counting the eggs is considered? As the method is being used this is not done in the lab but online by a technician from Techion Group Ltd.? This should be explained if not also considered with respect to the time calculations.

Reviewer #3: Y

Reviewer #4: Comments attached

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

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 article presented by the authors is relevant to the field of public health and provides useful information for STH deworming programs. Through a detailed analysis on the performance, time, and cost of three different FEC techniques they provide useful recommendations that can be readily applied. Therefore, I recommend publication of the article after a few minor revisions have been addressed.

Reviewer #2: This is a very important and potentially most relevant paper for all those interested in the monitoring of STH deworming programs. I find it very clearly presented and the data novel, convincing as well as significant.

Reviewer #3: (No Response)

Reviewer #4: Comments attached

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

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

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.

Attachments

Attachment

Submitted filename: Comments.docx

Dear Dr. Coffeng,

We are pleased to inform you that your manuscript 'A general framework to support cost-efficient fecal egg count methods and study design choices for large-scale STH deworming programs – monitoring of therapeutic drug efficacy as a case study' 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,

David Joseph Diemert, M.D.

Academic Editor

PLOS Neglected Tropical Diseases

Cinzia Cantacessi

Section Editor

PLOS Neglected Tropical Diseases

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