PONE-D-24-07797Novel Insights into Immune Stress Markers Associated with Myxosporeans Gill Infection in Nile Tilapia (Molecular And Immunohistochemical Studies)PLOS ONE

Dear Dr. Mohammed,

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Ishtiyaq Ahmad, Ph.D

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: 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 study conducted by Ramadan et al. is very interesting and will be useful for PLOS one readers. The manuscript the hypothesis of the study is clear and the manuscript is well-written. Minor issues are needed for improving the manuscript before being published.

Abstract

1-the abstract should be informative, all genes abbreviations should be mentioned in full formula for the first time, then use the abbreviated form

2- the authors should mention the methodology in brief (I mean, no of fish used in the study, no of groups, no of replicates, duration of the study).

3- The abbreviated form of genes should be italic

4-line 69, the abbreviations should be written in full formula

5-line 71, the abbreviated gene name should be italic

6-line 98, Oreochromis niloticus should replace by O. niloticus

7-line 153, µl should be replaced by µL and should be revised in all manuscript.

8-line 179, remove (,)

9-line 182, add the name of anti-coagulant

10-190, which organs were sampled and no of samples

11-line 202, what about normality test?

Reviewer #2: Dear Editor

I would like to thank you for giving me this opportunity to review this manuscript.

This manuscript aims to assess the stress response of Nile Tilapia to the infection by Myxosporeans via evaluating immune response at different levels, protein changes, hormonal and tissue histopathological and immunohistochemical alterations with good correlation of the results and interpretation of data based on the obtained results. This is an interesting study however the following points need to be addressed:

The title of the manuscript is very novel and defines the status of the article. All figures are beneficial in illustration of the results.

The authors mentioned that blood samples from five infected fish and five uninfected fish were subjected to biochemical analysis, so the authors before this step were examined the fish to determine whether they infected or not, so the handling of fish is considered a stressor that may alter the level of serum cortisol. So, you must highlight light after which period from handling you collect the blood samples?

line 183, the authors mentioned that: had blood drawn from their caudal veins. To extract serum by biochemical analysis,), please this sentence grammatically.

line 184, the authors mentioned that: the sample was then frozen at −20 °C.), you should mention that this was a serum sample.

In the histopathological section, why did the authors select TNF-α and iNOS to perform tissue expression?

the authors should mention the software used for image capturing and scale bar application.

the authors should clarify the significance in table (5).

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Reviewer #1: No

Reviewer #2: No

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Attachments

Attachment

Submitted filename: reviewer report for PLOS one 15-3-2024.docx

Novel Insights into Immune Stress Markers Associated with Myxosporeans Gill Infection in Nile Tilapia (Molecular And Immunohistochemical Studies)

PONE-D-24-07797R1

Dear Dr. Mohammed,

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,

Ishtiyaq Ahmad, Ph.D

Academic Editor

PLOS ONE

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

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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

**********

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

Reviewer #1: Yes

Reviewer #2: 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

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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 #2: 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: Congratulation. All comments have been addressed by authors. No additional comments are required. Now the manuscript is suitable for publication

Reviewer #2: All my inquiries have been thoroughly addressed by the authors of this manuscript and consequently I accept it for publication

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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

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