PONE-D-19-23901

Transfer of skin microbiota between two dissimilar autologous microenvironments: a pilot study

PLOS ONE

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PLOS ONE

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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: No

Reviewer #2: Yes

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

Reviewer #1: Yes

Reviewer #2: Yes

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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

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Reviewer #1: Yes

Reviewer #2: 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: The primary concern is a lack of agreeable definition for “transfer”. In this space the authors have assumed that the detection of DNA signature 0 or 24 hour after swabs constitutes transfer. As written, there is a possibility that they only transferred DNA, not viable organisms. In fact, the culture results in Table 3 argue strongly that they did not transfer a “viable community”, but at best transferred two coagulase negative Staph strains. Traditional culture techniques should not miss Staph epi – thus the lack of S. epi growth in the T24 cultures is concerning that no viable organisms were transferred in the first place.

A control could be performed swabbing the back with heat killed samples from the arm (i.e. culture Staph from the arm, heat kill it, then see if the bacterial 16S signature is present on the T0 and T24 samples). If the non-viable swab shows a less impressive transfer, the authors would have a much stronger claim. IE consider the direct non-viable culture swab a DNA-only, positive control to compare. An alternate approach would be to take the T0 swab and dip it in diluted EtOH prior to swabbing the back. If the 16S signature differed between the T24 and T24+EtOH group (versus a blank swab dipped in EtOH alone) the authors might have a stronger case that the T24 signature was from viable organisms.

Short of control samples, the entire paper (especially the abstract) requires that the language be amended to outline that they have transferred the DNA profile of the arm to the back. They have not provided the results needed to claim that they “can move viable skin organisms from one site to another”. There is no shame in showing that they were the first to transfer a partial DNA signature, but need to do further work to establish if they transferred viable organisms. But this paper claims multiple times to have transferred viable communities when they do not demonstrate that with their data.

This also needs to be discussed in detail in the limitations section, rather than the glancing comment contained now. As constructed, I feel that the overall language is misleading as it implies that swabs could be used from person A to person B to transfer viable organisms. This may be true, and is indeed “worthy of further investigation”, but without demonstrating colonization (even for as little as 24 hours), I cannot endorse this manuscript.

Minor concern:

In Fig 5, the authors state that the orange symbols – “the community has shifted towards the Ba samples (red) and away from the Bb samples (yellow)”. I don’t see this claim supported at all, can they provide mathematical support for this?

Reviewer #2: Perin et al present skin microbiome transplantation auto-transplantation from a site on the back to an arm site study in four patients. This work results interesting for the community but some aspects need to be clarified.

Major comments:

-It results unclear how it was measured the viability of the applied organisms on the transplant. It should be clarified how many live bacteria were transferred. Viability of the isolated microorganisms should be assayed and quantified

-Figures 3 and 5 should include the data of T24

Minor comments:

-What is the composition of the saline solution used for recovery?

-Tables are very hard to read. Version with improved resolution should be provided.

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Reviewer #1: No

Reviewer #2: No

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Transfer of skin microbiota between two dissimilar autologous microenvironments: a pilot study

PONE-D-19-23901R1

Dear Dr. Perin,

We are pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it complies with all outstanding technical requirements.

Within one week, you will receive an e-mail containing information on the amendments required prior to publication. When all required modifications have been addressed, you will receive a formal acceptance letter and your manuscript will proceed to our production department and be scheduled for publication.

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With kind regards,

Miroslav Blumenberg, PhD

Academic Editor

PLOS ONE

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

Reviewer #2: All comments have been addressed

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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 #1: Yes

Reviewer #2: Yes

**********

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

Reviewer #1: Yes

Reviewer #2: 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 #1: Yes

Reviewer #2: Yes

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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 #1: Yes

Reviewer #2: 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 #1: All of my issues have been addressed. I think the paper is sound and will be of use for the research community - with the admitted limitations added I think they present their data in a fair light (showing the promise without over stating).

Reviewer #2: Authors addressed all the relevant issues pointed during review. Missing data from graphics has been clarified and Tables improved.

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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.

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