PONE-D-22-17357Novel strains of Campylobacter cause diarrheal outbreak in Rhesus macaques (Macaca mulatta) of Kathmandu ValleyPLOS ONE

Dear Dr. Karmacharya,

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Dwij Raj Bhatta, PhD

Academic Editor

PLOS ONE

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Additional Editor Comments:

This important research on Campylobacter spp. and role in epidemic diarrhoea in Primates needs minor revision! The authors are suggested to answer queries of reviewers& adress comments from editor! In the abstract section highlighted text is confusing, sentence be changed! commensal flora of which animal species ??chicken?

In introduction part:mention types of standard cultural methods and biochemical identification ,confirmation of these microaerophilic bacteria briefly with proper reference ! Whether the molecular methods applied in the present research is enough to claim novel species? Why cultural methods not applied first to identify isolates and followed by Sequencing of such isolates?

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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: I Don't Know

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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: This study reports the novel strains of Campylobacter causing diarrhoeal outbreaks in monkeys in two holy sites of Kathmandu valley.

Major comments:

1.In abstract, the authors have mentioned “Opportunistic diarrheal samples of monkeys were analyzed to ………………” It is better to write diarrhoeal samples only instead of opportunistic samples.

2.In abstract, your result highlights the detection of Campylobacter in soil and water samples but methodology part is missing. Please mention about collection and processing of soil and water samples

3. In methods section, line 99, the authors have mentioned water samples were collected form ponds. In contrast to this, in line 102, they have mentioned 5 water samples from river and drinking water sources. Please write exactly which samples you have collected. Also elaborate on the collection and transportation of water samples.

4.In figure 1, please show the sample collection sites for water and soil samples

Minor comments:

Please correct the grammatical errors in the text such as

1.Line 81: Please rearrange the sentence.

2.Line 100: Please mention the name of the laboratory instead of our lab.

3.Line 111: Please write the symbol µ instead of u

4.Line 111-113: Please complete the sentence “ The cycling…….4°C.”

Reviewer #2: The manuscript is well written. Its a novel finding in Nepal and authors have used adequate advanced techniques for the laboratory experiment. However, only animal and environmental samples are included and human samples are not included in this study. Therefore the role of novel strain of the organism in human infections needs further research work with inclusive of human samples. Language of the manuscript at same places needs revision.

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Reviewer #1: No

Reviewer #2: No

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Attachments

Attachment

Submitted filename: PONE-D-22-17357 (1).pdf

PONE-D-22-17357R1Novel strains of Campylobacter cause diarrheal outbreak in Rhesus macaques (Macaca mulatta) of Kathmandu ValleyPLOS ONE

Dear Dr. Karmacharya,

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 Feb 11 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,

Dwij Raj Bhatta, PhD

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:

1. Please add a paragraph in introduction section /elaborate, by mentioning name of all species of a genus Capylobacter as per official nomenclature &mention determinative morphological biochemical characteristics, host range and medical importance of all species

2. In introduction part: briefly mention types of standard cultural methods and biochemical identification, confirmation of these microaerophilic bacteria with proper reference!

​

3. Whether the molecular methods applied in the present research is enough to claim novel species? Why cultural methods not applied first to identify isolates and followed by Sequencing of such isolates

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

Reviewers' comments:

[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.

PONE-D-22-17357R2Novel strains of Campylobacter cause diarrheal outbreak in Rhesus macaques (Macaca mulatta) of Kathmandu ValleyPLOS ONE

Dear Dr. Karmacharya,

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 Feb 17 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,

Dwij Raj Bhatta, PhD

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):

Need to address specific comment from academic editor made on their revision 1 manuscript previously sent to authors !! Unfortunately they have submitted manuscript again as revision 2 but it does not satisfy editor's comments and questions previously sent!

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Reviewers' comments:

[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.

Novel strains of Campylobacter cause diarrheal outbreak in Rhesus macaques (Macaca mulatta) of Kathmandu Valley

PONE-D-22-17357R3

Dear Dr. Karmacharya,

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,

Dwij Raj Bhatta, PhD

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

The reaserch gives important information on campylobacter species that were causative agent of dirrhoea in monkeys in Nepal and confirmed by WGST ! Revision 3 manuscript with track change has been reviewed and rebuttal letter also viewed! ! Revision 3 of the manusctipt be published along with supporting informations and author explanations !

Reviewers' comments: