-->PONE-D-25-63093-->-->Carissa spinarum L. aqueous root extracts ameliorate oxidative stress-induced infertility in adult male wistar rats-->-->PLOS One

Dear Dr. Ouma,

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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

Reviewer #3: Yes

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-->2. Has the statistical analysis been performed appropriately and rigorously? -->

Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: 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

Reviewer #3: 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

Reviewer #3: 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 article entitled " Carissa spinarum L. aqueous root extracts ameliorate oxidative stress-induced

infertility in adult male wistar rats" is interesting and shall be revised before considering in this journlal for publication.

1. In the abstract, state which solvent extract is used for the invivo study.

2. In the methodology, clearly state in invivo study, about which solvent's extract have been studied,

3. The number of animals used is 5, justtify the sample calculation method or how the number is selected.

4. State the study followed ARRIVE guidelines along with EU guidelines

5. Skecth some of the major phytochemical compounds profiled by GCMS

6. Discuss the rationale of selecting this plant and extract for the specific study.

7. Conclusion shall be specific and mention the predicted hormonal influence and the steroidal activation related to the plant.

Reviewer #2: Carissa spinarum L. aqueous root extracts ameliorate oxidative stress-induced infertility in adult male Wistar rats’’ is a well-organized study. All the objectives of research are clear and well-designed. Minor corrections need to be considered.

•Some typing and grammatical errors were observed. Please correct it.

•Drugs and chemicals should be written separately.

•The statistical symbols (for comparison) within tables and figures are not defined in legends. Kindly include its details for clarity.

•How the doses were selected, is there any previous study?

Reviewer #3: This is a well-designed and highly valuable pharmacological study. Validating the traditional medicinal use of Carissa spinarum for managing male infertility is a great contribution to reproductive biology and natural product research.

I appreciate your comprehensive methodology. Using metronidazole to induce oxidative stress in the Wistar rat model provided a robust baseline, and your evaluation of both the phytochemical profiles and in vivo parameters (sperm characteristics, biochemical assays, and histology) gives a very complete picture. The clear efficacy demonstrated at the 500 mg/kg dose for improving sperm count and reducing lipid peroxidation is particularly compelling.

I have no changes to request. Congratulations on a rigorous and well-executed paper.

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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.

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Reviewer #1: No

Reviewer #2: No

Reviewer #3: Yes:  Ganesh Praneeth Roy Avula

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Attachments

Attachment

Submitted filename: PONE-D-25-63093_reviewer.pdf

Carissa spinarum L. aqueous root extracts ameliorate oxidative stress-induced infertility in adult male wistar rats

PONE-D-25-63093R1

Dear Dr. Martin,

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,

Saira Saeed Khan, Ph.D.

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 #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

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-->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 addressed the quries raised and amended in the manuscript. The manuscript shall be accepted as per the editorial board decision.

Reviewer #2: (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 #1: No

Reviewer #2: Yes:  Sadia K.

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