Dear Dr. Zhiyi Zuo,

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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We look forward to receiving your revised manuscript.

Kind regards,

Huzaifa Umar

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?

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

Reviewer #1: Yes

Reviewer #2: Yes

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4. Is the manuscript presented in an intelligible fashion and written in standard English??>

Reviewer #1: Yes

Reviewer #2: Yes

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Reviewer #1: The study provides valuable insights into the gut microbiota–immune system interaction in sleep deprivation but requires improvements in data interpretation, clarity of methodology, and discussion depth.

1. Refine the title for conciseness while maintaining key elements; for example, Sleep Deprivation Disrupts Gut Microbiota Rhythms and Inflammatory Responses in Mice.

2. Improve grammar and specificity and include quantitative results in the abstract section.

3. Condense background information on circadian rhythms in the introduction section.

4. Explicitly state hypothesis and expected outcomes in the introduction section.

5. Strengthen rationale for behavioral assessments.

6. Provide details on microbiota sequencing normalization methods.

7. Clarify whether experimenters were blinded in behavioral assessments.

8. Ensure consistent use of statistical markers in figures and tables.

9. Include correlation analysis between cytokines and microbiota shifts.

10. Behavioral results should include effect size.

11. Improve figure clarity and readability.

12. Some speculative claims lack direct experimental validation (e.g., gut microbiota driving anxiety).

13. The clinical implications should be emphasized—how does this relate to human sleep disorders?

14. Lack of mechanistic validation (e.g., microbiota transplant studies).

15. Provide stronger mechanistic links between microbiota and behavior.

16. Expand discussion on clinical relevance.

Reviewer #2: he study employs a robust experimental design with comprehensive temporal sampling, enabling a detailed analysis of circadian disruptions caused by sleep deprivation. The integration of microbiome sequencing with cytokine profiling provides compelling evidence of altered microbial and immune rhythms, contributing valuable knowledge to the field of chronobiology and neuroimmunology. The findings have potential implications for understanding the health consequences of sleep loss and could inform therapeutic strategies aimed at mitigating circadian and inflammatory disturbances. Given the novelty, methodological rigor, and relevance to human health, I recommend acceptance of this manuscript for publication.

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Reviewer #1: No

Reviewer #2: No

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Sleep deprivation disrupts diurnal rhythmicity of gut microbiota and blood inflammatory cytokines in mice

PONE-D-25-03292R1

Dear Dr. Zhiyi,

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 email 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,

Huzaifa Umar

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

Reviewers' comments: