PONE-D-23-13258The Investigation of Antibacterial Properties of Peptides and Protein Hydrolysates Derived from Serum of Asian Water Monitor (Varanus salvator)PLOS ONE

Dear Dr. Thanasak,

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.

Please submit your revised manuscript by Jul 29 2023 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,

Md. Imtaiyaz Hassan, Ph.D.

Academic Editor

PLOS ONE

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When submitting your revision, we need you to address these additional requirements.

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2. We noticed you have some minor occurrence of overlapping text with the following previous publication(s), which needs to be addressed:

https://www.mdpi.com/1422-0067/22/15/7959

https://www.frontiersin.org/articles/10.3389/fmicb.2021.771527/full

In your revision ensure you cite all your sources (including your own works), and quote or rephrase any duplicated text outside the methods section. Further consideration is dependent on these concerns being addressed.

3. We note that the grant information you provided in the ‘Funding Information’ and ‘Financial Disclosure’ sections do not match. 

When you resubmit, please ensure that you provide the correct grant numbers for the awards you received for your study in the ‘Funding Information’ section.

4. We note that you have stated that you will provide repository information for your data at acceptance. Should your manuscript be accepted for publication, we will hold it until you provide the relevant accession numbers or DOIs necessary to access your data. If you wish to make changes to your Data Availability statement, please describe these changes in your cover letter and we will update your Data Availability statement to reflect the information you provide.

Additional Editor Comments:

A critical assessment of the present knowledge with some clear conclusions what all these results mean, and directions for future research and potential applications should be strengthened.

I found minor issue, many typos and grammatical errors are seen in the paper. There are grammatical mistakes and typographical errors in the manuscript. The author should recheck this manuscript carefully and remove all such errors.

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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: 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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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: 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 authors demonstrated that "antimicrobial peptides were predicted which shows highly hydrophobic". It will be interesting to test and check their amyloidogenicity according the last review for AMPs - Int J Mol Sci. 2022 May 13;23(10):5463. doi: 10.3390/ijms23105463.

Line 45. “ceropin” correct to “cecropin”.

Lines 59-60. “Moreover, BmKn-2, an animal peptide especially extracted from scorpion venom, has shown the inhibitory effect on bacterial growth [11, 14-16].” I think that BmKn-2 is not discussed in the cited work #15. Please check.

Line 101. “digestive System” correct to “digestive system”.

Line 117. “a plain blood collecting tube” correct to “a plain blood collection tube”.

Lines 156-157. “The inhibitory percentage was calculated from [(OD control − OD peptide test)/OD control] × 100”. OD control – is it OD for positive or negative control? If this is the optical density for the negative control, then Table 1 should indicate the antimicrobial efficacy of the crude peptides and protein hydrolysates in comparison with the antibiotic, colistin.

Reviewer #2: The manuscript titled as “The Investigation of Antibacterial Properties of Peptides and Protein Hydrolysates Derived from Serum of Asian Water Monitor (Varanus salvator)” represents an interesting study and is well written that only need minor modifications. However, I feel that some relevant images can be added to make the study more attractive and interesting. Please consider the following points:

1. Title and abstract:

Title and abstract are accurate and adequately addressed. However, a graphical abstract can be added.

2. Introduction:

1. L- 42: Please give the examples of resistant and non-resistant pathogenic bacteria.

2. L-73: ‘evidence of reduced in bacterial growth’ these sentence needs to be re-written.

3. L-80: ‘according to the list of Thailand protected species’, please give an appropriate reference for this sentence.

3. Materials and Methods

1. L-27: Why Lowry's method is selected for protein determination over other methods?

2. L-130: Why boiling for 10 minutes is done?

3. L-155: ‘Colistin was used as positive control while hydrolysate buffer was used as negative control’. Please specify reason for this statement.

4. L-156: mention complete formula in mathematical manner.

5. L-169: Specify the reason for the use of nitrogen as a drying gas.

4. Results:

1. Table 2. Antimicrobial peptide prediction of Varanus salvator serum, using’ this line seems incomplete.

2. L-247 ‘5 Gram-negative and 2 Gram-positive bacteria,’ please mention the names of concerned bacteria and also mention reference for this statement.

3. L-269: ‘high isoelectric point’ please mention the numerical value also.

4. L-273 ‘some mechanisms can be proposed’ give brief description of the mechanisms that can be proposed.

5. Conclusion:

1. L-294 Antimicrobial peptides…..Hydrophobic, this line is not in flow with this paragraph, so needs to be re-written.

2. Conclusion section is written in brief. Please add more relevant points in it.

3. Also write a concluding line in the end with future scope of this research.

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Reviewer #1: No

Reviewer #2: No

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Attachments

Attachment

Submitted filename: Comments and review of paper[535].docx

PONE-D-23-13258R1The Investigation of Antibacterial Properties of Peptides and Protein Hydrolysates Derived from Serum of Asian Water Monitor (Varanus salvator)PLOS ONE

Dear Dr. Thanasak,

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.

Please submit your revised manuscript by Oct 06 2023 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,

Md. Imtaiyaz Hassan, Ph.D.

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:

Authors must address all the comments.

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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: (No Response)

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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: (No Response)

**********

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: (No Response)

**********

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: The authors did not answer on my previous question: "The authors demonstrated that "antimicrobial peptides were predicted which shows highly

hydrophobic". It will be interesting to test and check their amyloidogenicity according the last

review for AMPs - Int J Mol Sci. 2022 May 13;23(10):5463. doi: 10.3390/ijms23105463."

**********

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

**********

[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.

The Investigation of Antibacterial Properties of Peptides and Protein Hydrolysates Derived from Serum of Asian Water Monitor (Varanus salvator)

PONE-D-23-13258R2

Dear Dr. Thanasak,

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,

Md. Imtaiyaz Hassan, Ph.D.

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

The revised submission is acceptable now.

Reviewers' comments: