PWAT-D-21-00006

Operationalizing a routine wastewater monitoring laboratory for SARS-CoV-2

PLOS Water

Dear Dr. Kantor,

Thank you for submitting your manuscript to PLOS Water. After careful consideration, we feel that it has merit but does not fully meet PLOS Water'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 30 days. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at water@plos.org. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pwat/ 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 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'.

Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter.

We look forward to receiving your revised manuscript.

Kind regards,

Silvia Monteiro

Academic Editor

PLOS Water

Journal Requirements:

1. We ask that a manuscript source file is provided at Revision. Please upload your manuscript file as a .doc, .docx, .rtf or .tex. If you are providing a .tex file, please upload it under the item type ‘LaTeX Source File’ and leave your .pdf version as the item type ‘Manuscript’.

2. Please provide separate figure files in .tif or .eps format only, and remove any figures embedded in your manuscript file.  If you are using LaTeX, you do not need to remove embedded figures.

For more information about figure files please see our guidelines: 

https://journals.plos.org/water/s/figures

3. Please update the completed 'Competing Interests' statement, including any COIs declared by your co-authors. If you have no competing interests to declare, please state "The authors have declared that no competing interests exist". Otherwise please declare all competing interests beginning with the statement "I have read the journal's policy and the authors of this manuscript have the following competing interests:"

4. We notice that your supplementary figures are uploaded with the file type 'Manuscript' and are therefore included in the PDF. Please amend the file type to 'Supporting Information'. Please ensure that all Supporting Information files are included correctly and that each one has a legend listed in the manuscript after the references list.

5. In the online submission form, you indicated that "Data are available in the supplementary tables. Raw data for Figures 2 and 3 are available upon request." All PLOS journals now require all data underlying the findings described in their manuscript to be freely available to other researchers, either 1. In a public repository, 2. Within the manuscript itself, or 3. Uploaded as supplementary information.

This policy applies to all data except where public deposition would breach compliance with the protocol approved by your research ethics board. If your data cannot be made publicly available for ethical or legal reasons (e.g., public availability would compromise patient privacy), please explain your reasons by return email and your exemption request will be escalated to the editor for approval. Your exemption request will be handled independently and will not hold up the peer review process, but will need to be resolved should your manuscript be accepted for publication. One of the Editorial team will then be in touch if there are any issues.

6. Please amend your detailed Financial Disclosure statement. This is published with the article, therefore should be completed in full sentences and contain the exact wording you wish to be published.

i) Please include all sources of funding (financial or material support) for your study. List the grants (with grant number) or organizations (with url) that supported your study, including funding received from your institution. 

ii). State the initials, alongside each funding source, of each author to receive each grant.

iii). State what role the funders took in the study. If the funders had no role in your study, please state: “The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.”

iv). If any authors received a salary from any of your funders, please state which authors and which funders.

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Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. Does this manuscript meet PLOS Water’s publication criteria? Is the manuscript technically sound, and do the data support the conclusions? The manuscript must describe methodologically and ethically rigorous research with conclusions that are appropriately drawn based on the data presented.

Reviewer #1: Yes

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2. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: No

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3. Have the authors made all data underlying the findings in their manuscript fully available (please refer to the Data Availability Statement at the start of the manuscript PDF file)?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception. 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: No

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4. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS Water 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

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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 manuscript presents a coordinated analytical sequence for quantifying SARS-CoV-2 genome concentration in wastewater, which includes the steps of sample collection, laboratory testing and data analysis with quality assurance procedures. This information is worth sharing with readers, but the following issues need to be addressed before publication.

The first issue is about the substitution of non-detects with the half value of detection limit (line 312, Figure S1). I cannot understand why this substitution is necessary. When viral concentration is used for further analysis, such as a correlation analysis with prevalence level in a sewershed, this substitution distorts the statistical analysis. Please justify this appropriately if mandatory for authors.

The second issue is about the decision of positive/negative for samples with very low concentration of viral gene. Authors use three tubes per sample (page 7, line 118). If the reviewer does not miss it, there is no description how authors treat the result when only one or two tubes are positive. Please clearly specify in the text how to deal with this situation.

The third issue is about the data presentation in Figure 2. I’m not quite sure how to justify the usage of lowess-smoothed fitting function. This fitting must affect subsequent analysis, and I do not think this is well justified.

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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.

Do you want your identity to be public for this peer review? If you choose “no”, your identity will remain anonymous but your review may still be made public.

For information about this choice, including consent withdrawal, please see our Privacy Policy.

Reviewer #1: Yes: Daisuke Sano

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[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.

Operationalizing a routine wastewater monitoring laboratory for SARS-CoV-2

PWAT-D-21-00006R1

Dear Dr. Kantor,

We're pleased to inform you that your manuscript has been judged scientifically suitable for publication in PLOS Water. The academic editor and reviewers were very positive about your work and we sincerely appreciate your support of the journal and Open Science.

As a new journal, we are finalising our publishing workflow, so we appreciate your patience as your manuscript moves through the system. Soon, you'll receive an e-mail detailing the required amendments. When these have been addressed, you'll receive a formal acceptance letter. An invoice for payment, along with detailed instructions, will follow shortly after the formal acceptance. 

PLOS Water will formally begin publishing in early 2022 and your manuscript will be scheduled for publication as soon as possible after this point.To minimise any delay in the dissemination of this work, we encourage you to update the preprint that you have already posted with the most up-to-date version.

 

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, and 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 water@plos.org.

Thank you for your submission and trust in us as we launch this important new space for the study of water for people and the planet. We are thrilled that you are among the initial cohort of papers that is working to address the most pressing issues in water sanitation and water resources that are facing our world today.

Kind regards,

Silvia Monteiro

Academic Editor

PLOS Water

Additional Editor Comments (optional):

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation.

Reviewer #1: All comments have been addressed

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2. Does this manuscript meet PLOS Water’s publication criteria? Is the manuscript technically sound, and do the data support the conclusions? The manuscript must describe methodologically and ethically rigorous research with conclusions that are appropriately drawn based on the data presented.

Reviewer #1: Yes

**********

3. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: Yes

**********

4. Have the authors made all data underlying the findings in their manuscript fully available (please refer to the Data Availability Statement at the start of the manuscript PDF file)?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception. 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

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5. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS Water 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

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6. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: (No Response)

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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.

Do you want your identity to be public for this peer review? If you choose “no”, your identity will remain anonymous but your review may still be made public.

For information about this choice, including consent withdrawal, please see our Privacy Policy.

Reviewer #1: Yes: Daisuke Sano

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