PONE-D-21-24028Evaluation of SARS-CoV-2 in semen, seminal plasma, and final fraction with spermatozoa of COVID-19 patients at acute stage of infectionPLOS ONE

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Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: Yes

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

Reviewer #1: Yes

Reviewer #2: N/A

Reviewer #3: Yes

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Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: Yes

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Reviewer #1: No

Reviewer #2: Yes

Reviewer #3: Yes

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Reviewer #1: Reviewer’s report

Title: Evaluation of SARS-CoV-2 in semen, seminal plasma, and final fraction with spermatozoa of COVID-19 patients at acute stage of infection

Date: 31 August 2021

Manuscript ID: PONE-D-21-24028

Reviewer’s report:

In the manuscript titled “: Evaluation of SARS-CoV-2 in semen, seminal plasma, and final fraction with spermatozoa of COVID-19 patients at acute stage of infection” the authors evaluated the presence of SARS-CoV-2 in semen, seminal plasma and pallet samples of COVID-19 positive patients in the acute phase of infection. The topic could be of interest however several concerns need to be raised:

1- I suggest changing the “native semen” to semen which is more common.

2- Do the authors mean pallet after spinning the semen sample by “final fraction containing spermatozoa”? If yes change it to pallet.

3- Please write down these expressions: ESHRE, IFFS, ASRM in the instruction.

4- Introduction is too long. I recommend shortening it.

5- Please clarify and explain which criteria did the authors considered as an acute phase and moderate symptom?

6- Discussion lacks depth (it is purely descriptive).

7- It is noted that this manuscript needs a professional technical English editing service paying particular attention to English grammar and sentence structure.

Reviewer #2: The article from L. Delaroche et al. addresses the question of the presence of SARS-Cov-2 in semen from COVID-19 patients in the acute stage of infection. This study is an observational study.

Thirty-two men were included in the study and the virus was detected by RT-qPCR using the RealStar RT-PCR kit in native semen, seminal plasma and 80% fraction obtained after centrifugation of the semen on discontinuous density gradients. Only one volunteer presented positive SARS-Cov-2 RT-PCR in semen and seminal plasma, 80%fraction were negative. Following bacterial investigation of semen samples, the authors suggested that the SARS-Cov-2 presence in semen from one volunteer is more a consequence of oral contamination than e semen SARS-Cov-2 infection.

I have several comments:

I did not understand the reasons of semen home collection in 3 patients. The authors explain this: “according to the patient’s clinical condition”. This point needs to be clarified as 2 patients were symptomatic and the last was asymptomatic.

The semen investigation methods are not totally comprehensives. Linea 261: is the supernatant obtained from native semen centrifugation or from density gradient centrifugation? If is the first case, I suggest to present before “the remaining semen samples…(linea 258).

Table 1 and 2 reported the result characteristics in detail. For the reader, I think that table 1 could be more synthetic.

One patient had a positive semen. The patient performed semen probe at home and the precise bacterial study suggest a possible contamination from oropharyngeal secretions. In discussion, the authors mentioned the “The bacteria found can neither confirm nor strictly rule out a weak contamination of the semen sample by oropharyngeal secretions”. Indeed, higher concentrations of bacterial DNA were found in this specimen than in others but we did not know the levels which allows to conclude oral contamination in one and no in others. The sentence in abstract “the bacteria identified do not clearly rule out contamination by oropharyngeal secretions” could be modified according to “neither confirm nor rule out contamination” according to discussion.

Reviewer #3: A study with a final sample size of 32 COVID-19 positive volunteers aimed to determine whether SARS-CoV-2 could be detected in native semen, seminal plasma, and final fraction with spermatozoa samples. SARS-CoV-2 was detected in only one volunteer’s native semen and seminal plasma samples. The results are primarily descriptive.

Minor revisions:

1- Line 104: Indicate the type of summary statistics provided for age.

2- Patient characteristics: In addition to the frequencies, state the corresponding percentages.

3- Line 186: Provide the corresponding percentage and a 95% confidence interval for the 31/32 result.

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Reviewer #1: Yes: kajal Khodamoradi

Reviewer #2: Yes: Bujan Louis

Reviewer #3: No

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Evaluation of SARS-CoV-2 in semen, seminal plasma, and spermatozoa pellet of COVID-19 patients in the acute stage of infection

PONE-D-21-24028R1

Dear Dr. DELAROCHE,

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.

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Kind regards,

Pierre Roques, Ph.D.

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 #3: 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 #3: (No Response)

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

Reviewer #1: Yes

Reviewer #3: (No Response)

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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 #3: (No Response)

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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 #3: (No Response)

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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)

Reviewer #3: (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.

If you choose “no”, your identity will remain anonymous but your review may still be made public.

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Reviewer #1: Yes: kajal khodamoradi

Reviewer #3: No