Peer Review History

Original SubmissionJanuary 31, 2023
Decision Letter - Nabi Jomehzadeh, Editor

PONE-D-23-02752Assessment of four in vitro phenotypic biofilm detection methods and antimicrobial resistance in aerobic clinical bacterial isolatesPLOS ONE

Dear Dr. Tamang,

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 May 13 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,

Nabi Jomehzadeh, Ph.D (Assistant Professor)

Academic Editor

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: 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: 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: 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 author has tried to evaluate the resistance pattern of isolated bacteria in study period and has compared different methods for biofilm formation. Following comments need to be addressed appropriately:

1. Author should clearly mention the aim of the study in abstract and in main manuscript. Presently, the aim defined at two sites is different from each other.

2. In materials and methods, please mention clearly how was the susceptibility testing done for colistin and vancomycin? As per CLSI guidelines, Kirby Bauer method can't be used for these drugs.

3. In results: line 247, the total of samples given is 205, not 194 as mentioned by author.

4. In line 249, Please expand the term ACB because it is written first time here.

5. In table 2; what is the relevance of calculating p-values?

6. Table 6 is not needed. Author has already shown relationship between biofilm formation and drug resistance (MDR & XDR) in table 5.

Reviewer #2: Dear Authors. You are well done.

All the revisions and very important notes are documented on the text. To follow and track and complete your manuscript. It is need to update the references and adding as we mentioned in the text

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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.

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

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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.

Attachments
Attachment
Submitted filename: PONE-D-23-02752_reviewer.pdf
Attachment
Submitted filename: 2nd assignment-PONE-D-23-02752 after revision.pdf
Revision 1

General comments (Journal requirements)

1. Please ensure that your manuscript meets PLOS ONE's style requirements, including those for file naming. The PLOS ONE style templates can be found at

https://journals.plos.org/plosone/s/file?id=wjVg/PLOSOne_formatting_sample_main_body.pdf and

https://journals.plos.org/plosone/s/file?id=ba62/PLOSOne_formatting_sample_title_authors_affiliations.pdf

Response: We thank you for this useful suggestion. We followed the links and made the necessary changes to the manuscript. As suggested, upon improvements, we have prepared and named three different files as 'Response to Reviewers' (for rebuttal letter), and ' Manuscript with Track Changes’ (for a marked-up copy of the manuscript), and ‘Revised Manuscript’ (for revised paper). Additionally, we have checked and attest that all formatting and style requirements have been met in the manuscript.

2. Please upload a completed version of your questionnaire as Supporting Information when you resubmit your manuscript.

Response: We have uploaded a completed version of questionnaire as “Supporting Information”.

3. Please ensure that you have an ORCID iD and that it is validated in Editorial Manager.

Response: Validated ORCID iD has been added.

4. Please include your full ethics statement in the ‘Methods’ section of your manuscript file. In your statement, please include the full name of the IRB or ethics committee who approved or waived your study, as well as whether or not you obtained informed written or verbal consent. If consent was waived for your study, please include this information in your statement as well.

Response: We have now added a full ethics statement in the ‘Methods’ section of the manuscript file, as well as a sentence concerning the written informed consent.

Response to Reviewer 1

1. Author should clearly mention the aim of the study in abstract and in main manuscript. Presently, the aim defined at two sites is different from each other.

Response: First of all, we will like to express our sincere gratitude for your kind suggestions. We appreciate the time and effort that you dedicated to providing feedback on our manuscript and are grateful for the insightful comments on our paper.

Thank you for pointing this out. We agree with this comment. Therefore, following the reviewer suggestions, the aim of the study throughout the manuscript (abstract and main text) has been updated as ... to evaluate diagnostic parameters of four in vitro phenotypic biofilm detection assays in relation to antimicrobial resistance in aerobic clinical bacterial isolates...

2. How was the susceptibility done for colistin and vancomycin? As per CLSI guidelines, Kirby Bauer method can't be used for these drugs.

Response: Thank you for pointing this out. We have now studied the CLSI guideline (32th edition), and as is explicitly stated in it, we have as of this this time amend this error by erasing the susceptibility testing for colistin and vancomycin. Further, we will implement this information in routine diagnosis too.

3. Total is not 194. It is 205. Please confirm.

Response: Dear reviewer, there were 194 samples that tested culture positive. Of these 162 samples yielded monobacterial growth, while 32 samples yielded 2 bacterial growth

162 samples = 162 isolates

32 samples = 64 isolates

---------------------------------------------------------------

194 total samples = 226 total isolates

Therefore, we confirm total samples as 194.

4. Expand ACB.

Response: Thank you for pointing this error. We have expanded the abbreviation accordingly. … Acinetobacter calcoaceticus-baumanii (ACB) complex….

5. What is the relevance of calculating p-value here in this table?

Response: Thank you for this comment. Actually, the table shows interesting and significant association of pathogen with the samples. For e.g., as we know, E. coli as a significant uropathogen, S. Typhi as a significant bloodstream pathogen, non-fermenters as the significant respiratory pathogens, and lastly S. aureus as the significant wound pathogen; all of these routine findings are explicitly portrayed in the table (significant correlation as per p-value reiterates the preexisting fact).

6. How do you differentiate between colonizers and pathogens?

Response: As we know, colonization is the presence of bacteria on a body surface (like on the skin, mouth, intestines or airway) without causing disease in the person. Infection is the invasion of a host organism's bodily tissues by disease-causing organisms. The samples obtained in the study were stringently from hospital visiting patients clinically suspected of infection. Hence, they were most likely to be pathogens.

7. Table 6 is not needed. Author has already shown relationship between biofilm formation and drug resistance (MDR & XDR) in table 5.

Response: Dear Reviewer, table 5 is discrete to table 6 in context to simply portraying the pathogen specific incidences of MDR-, XDR- or biofilm formation. The latter distinctly, however, shows antibiotic specific incidences of resistance (%) to each of the tested, be it with regard to MDR-, XDR-, or biofilm-MDR-, or biofilm-XDR-isolates. Furthermore, table 6 summarizes the overall resistance to each antibiotic tested, while table 5 fundamentally shows the frequencies. Therefore, we sincerely request you to reconsider the urgency of table 6.

Response to Reviewer 2

1. All the revisions and very important notes are documented on the text. To follow and track and complete your manuscript. It is need to update the references and adding as we mentioned in the text

Thank you for your kind suggestions. We have now updated the title as “Assessment of four in vitro phenotypic biofilm detection methods in relation to antimicrobial resistance in aerobic clinical bacterial isolates” We have added all the mentioned references in the study as well as the indicated change(s).

Attachments
Attachment
Submitted filename: Response to Reviewers.docx
Decision Letter - Nabi Jomehzadeh, Editor

PONE-D-23-02752R1Assessment of four in vitro phenotypic biofilm detection methods in relation to antimicrobial resistance in aerobic clinical bacterial isolatesPLOS ONE

Dear Dr. Tamang,

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 Nov 27 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,

Nabi Jomehzadeh, Ph.D (Assistant Professor)

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 (if provided):

"We note that one or more reviewers has recommended that you cite specific previously published works. As always, we recommend that you please review and evaluate the requested works to determine whether they are relevant and should be cited. It is not a requirement to cite these works. We appreciate your attention to this request.”

[Note: HTML markup is below. Please do not edit.]

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

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 #2: Yes

Reviewer #3: Yes

**********

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

Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: 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

Reviewer #3: 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

Reviewer #2: Yes

Reviewer #3: 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: the author has addressed all the comments and has made appropriate changes. The author has provided the data in well-understandable manner and has included the tables appropriately.

Reviewer #2: Dear Author

You are doing well

There are simple errors in writing the two references in numbers 13 and 41 in the list of references of the revised manuscript which the contributing authors are missing. Thus, the below the revisions:-

1-Reference 13:- The corrected and revised form of reference 13 is as follow:-Al-Ouqaili, M.T.S., Al-Kubaisy, S.H.M., Al-Ani, N.F.I. Biofilm antimicrobial susceptibility pattern for selected antimicrobial agents against planktonic and sessile cells of clinical isolates of staphylococci using MICs, BICs, and MBECs. Asian Journal of Pharmaceutics, Volume 12, Issue 4, October-December 2018, Pages S1375-S1383 instead of the citation form in the list of references.

2-Reference 14:- The corrected and revised form of reference 41 is as follow:-Al-Ouqaili, MTS, Al-Taei, SA, Al-Najjar A. Molecular Detection of Medically Important Carbapenemases Genes Expressed by Metallo-β-lactamase Producer Isolates of Pseudomonas aeruginosa and Klebsiella pneumoniae. Asian Journal of Pharmaceutics • Jul -Sep 2018 (Suppl ) • 12 (3) | S991 instead of the citation form in the list of references.

Reviewer #3: The authors have well addressed the reviewer’s comments however there are minor clarifications in results required from authors.

Table 2: why no of isolates are more than no of samples of urine blood and sputum

Table 2: what is the significance of p-value

Table 3: what is the significance of bold figures. e.g., in S. Typhi ceftazidine in bold with 3 resistant and not ciprofloxacin etc with 6 resistant isolates. Similar is the case with S. aureus and P. aeruginosa.

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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

Reviewer #3: Yes: Abu Baker Siddique

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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.

Attachments
Attachment
Submitted filename: Reviewer Comments.docx
Revision 2

We are grateful to the reviewers for their insightful comments on this study. We have been able to incorporate changes to reflect most of the suggestions provided by the reviewers.

Attachments
Attachment
Submitted filename: Response to Reviewers.docx
Decision Letter - Nabi Jomehzadeh, Editor

Assessment of four in vitro phenotypic biofilm detection methods in relation to antimicrobial resistance in aerobic clinical bacterial isolates

PONE-D-23-02752R2

Dear Dr. Tamang,

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.

An invoice for payment will follow shortly after the formal acceptance. To ensure an efficient process, please log into Editorial Manager at http://www.editorialmanager.com/pone/, click the 'Update My Information' link at the top of the page, and double check that your user information is up-to-date. If you have any billing related questions, please contact our Author Billing department directly at authorbilling@plos.org.

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,

Nabi Jomehzadeh, Ph.D (Assistant Professor)

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 #3: All comments have been addressed

**********

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 #3: Yes

**********

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

Reviewer #3: 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 #3: 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 #3: 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 #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.

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 #3: No

**********

Formally Accepted
Acceptance Letter - Nabi Jomehzadeh, Editor

PONE-D-23-02752R2

Assessment of four in vitro phenotypic biofilm detection methods in relation to antimicrobial resistance in aerobic clinical bacterial isolates

Dear Dr. Tamang:

I'm pleased to inform you that your manuscript has been deemed suitable for publication in PLOS ONE. Congratulations! Your manuscript is now with our production department.

If your institution or institutions have a press office, please let them know about your upcoming paper now to help maximize its impact. If they'll be preparing press materials, please inform our press team within the next 48 hours. 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.

If we can help with anything else, please email us at plosone@plos.org.

Thank you for submitting your work to PLOS ONE and supporting open access.

Kind regards,

PLOS ONE Editorial Office Staff

on behalf of

Dr. Nabi Jomehzadeh

Academic Editor

PLOS ONE

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