PONE-D-23-38195Microbial contamination of spittoons and germicidal effect of irradiation with Krypton Chloride excimer lamps (Far UV-C 222 nm)PLOS ONE

Dear Dr. TANIMOTO,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’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.

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ACADEMIC EDITOR: Please kindly attend to comments below. Thank you

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Please submit your revised manuscript by Mar 20 2024 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file.

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We look forward to receiving your revised manuscript.

Kind regards,

Charles Odilichukwu R. Okpala

Academic Editor

PLOS ONE

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5. Please amend the manuscript submission data (via Edit Submission) to include author Yoshimasa Ogawab, Takayuki Nambuc, Toru Koib, Toshinori Okinagac and Kazuyo Yamamotoa.

Additional Editor Comments:

Please, authors kindly attend to the reviewers' comments

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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: Yes

Reviewer #2: Partly

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

Reviewer #1: Yes

Reviewer #2: I Don't Know

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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: No

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4. 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: No

Reviewer #2: No

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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: a. The study and findings are quite interesting. However, the paper would be easier to read if both the Methodology and Results are arranged in subsections; for example, the methodology could have subsections such as Collection of samples, determination of microbial load, identification of contaminants etc.

b. There is a question in the methodology section that needs to be answered.

Reviewer #2: Title: Microbial contamination of spittoons and germicidal effect of irradiation with Krypton Chloride excimer lamps (Far UV-C 222 nm)

Comments to the authors

I appreciate the author's hard effort.

The authors measure the disinfectant effect of 222 nm Far UV-C radiation on dental spittoon, one may determine the bioburden of the spittoon and assess the sensitivity of two oral Streptococcus and two Enterococci to the radiation.

Although the study presented here is good, several crucial data points are missing.

Major concern

1. The main issue is that each method is included in a single paragraph. It ought to be divided into multiple sections. Each technique must to have a unique the title. Every protocol should be written in a single, concise sentence. Every protocol should be thoroughly written

2. Did you create a growth curve for line 105, "First, each strain was grown to the mid-logarithmic growth phase"? How did you find out each strain's mid-logarithmic development phase?

3. The 16s rRNA sequencing technique is absent. This is a significant experiment; could you further elaborate on the 16s rRNA sequencing methodology?

4. The manuscript needs to include the steps for phylogenetic diversity, OTU grouping and taxonomical assignment, trimmed sequence, and data sequence filtering in QIIME 2.

5. How did you determine that the change is significant? The paper has to include alpha and beta diversity analysis along with figures. How many germs are extracted from the sample and how many are common? There should be a Venn diagram.

Minor Comments

- Please add reference to line 43-44, 50- 51

- Italicize the bacterial name as in line 139 “Pseudomonas,” Pls check the whole manuscript.

- Line 84, change “was attempted” to “ was conducted”

- Line 93, what is the size of amplified fragment? Add primers sequences used?

- Line 106, “and suspended in PBS? Is not clear, GAM broth or PBS?. Write the whole protocol in details.

- - Lines 107–109 This procedure is quite brief. "How did you perform the irradiation? Could you perhaps elaborate on the protocol in a separate paragraph?

- - Lines 109–110 The CFU method should be described in detail as well.

- Line 115 “Our results suggest that 222 nm Far UV-C irradiation may inhibit the pathway of bacterial transmission by droplets from sink surfaces.” Should be moved to result.

- Line 130, “number of bacteria” Do you mean CFU? Why did you count 300 colonies when there were actually more?

- Line 141-142 “Ten of these species were present in three samples with more than 0.5% in common” Is it among the 15 sample or among the whole species numbers? You need to explain or justify why it precent in very low percentage.

- Enter the complete names of the SD, SE, and IC below the Table 1 as short notes. Include the "CFU" in the table itself as well.

-A native English speaker should review the paper. Please review the entire document.

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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: Yes: Ifeoma Maureen Ezeonu, Ph.D.

Reviewer #2: Yes: Anis Rageh Al-Maleki

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PONE-D-23-38195R1Microbial contamination of spittoons and germicidal effect of irradiation with Krypton Chloride excimer lamps (Far UV-C 222 nm)PLOS ONE

Dear Dr. TANIMOTO,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’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.

==============================

ACADEMIC EDITOR: Please kindly see comments below.==============================

Please submit your revised manuscript by Jul 25 2024 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ 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 academic 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'.

If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter.

If applicable, we recommend that you deposit your laboratory protocols in protocols.io to enhance the reproducibility of your results. Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: https://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols. Additionally, PLOS ONE offers an option for publishing peer-reviewed Lab Protocol articles, which describe protocols hosted on protocols.io. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols.

We look forward to receiving your revised manuscript.

Kind regards,

Charles Odilichukwu R. Okpala

Academic Editor

PLOS ONE

Journal Requirements:

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.

Additional Editor Comments:

I have critically examined the revised manuscript, and authors have addressed critical comments of the work.

Authors, kindly attend to the following and make sure to only upload the final draft. Use red font to indicate the revised areas.

a) Kindly include in your introduction about the global ramifications of microbial contamination in clinical settings - mortality, etc, bring in published data showing different continents, narrow down to Asia, then one in Japan. (this should be your paragraph 1). Make sure to contextualize the paragraph 1 to the subject of this work.

b) In materials and methods, please separate out the data analysis aspects and create the last subsection "statistical analysis", and have all of them there.

c) Your discussion needs additional depth. Please, provide deeper explanation of the results, not just about 'what', but more about 'how' and 'why?' . Please also, kindly make sure you include (Refer to Table 1) (Refer to Figure 2) (Refer to Figure 3) (Refer to Figure 4) in all the places where data of each has been discussed. So, all the Tables/Figures in the results, must be captured in the discussion

The editor will thoroughly examine these in your revised manuscript. Thank you very much and look forward to your revised manuscript.

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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. 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

**********

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?

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

**********

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

**********

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: Although, one would have wished for the use of a larger sample size in this study, the paper can be accepted because of the detailed analysis of the few samples employed. Moreover, the paper introduces a novel method of reducing contamination and transmission of infections via reusable dental instruments.

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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: Yes: Ifeoma Maureen Ezeonu

**********

[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.

Microbial contamination of spittoons and germicidal effect of irradiation with Krypton Chloride excimer lamps (Far UV-C 222 nm)

PONE-D-23-38195R2

Dear Dr. TANIMOTO,

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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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 -- 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 onepress@plos.org.

Kind regards,

Charles Odilichukwu R. Okpala

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

I have critically evaluated the revised manuscript, as well as the responses reviewers provided, and very satisfied to accept it for publication.

Reviewers' comments: