PONE-D-24-36152Microbiota composition of Culex perexiguus mosquitoes during the West Nile virus outbreak in southern SpainPLOS ONE

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“This work was supported by the MICIU/AEI/10.13039/501100011033 under grant PID2020-118205GB-I00. Additional support for this study comes from the European Commission – NextGenerationEU (Regulation EU 2020/2094), through CSIC’s Global Health Platform (PTI Salud Global). M Garrido was supported by the P9 program for the Incorporation of Young Doctors funded by the University of Granada and granted by PID2022-137746NA-I00 funded by MICIU/AEI/10.13039/501100011033 and by ERDF/EU. Jesús Veiga received financial support from the Juan de la Cierva program (FJC2021-048057-I) and Marta Garrigós from the FPI program (PRE2021-098544) funded by MICIU/AEI/10.13039/501100011033 and the European Union NextGenerationEU/PRTR. MJRL was funded by the Agencia Estatal de Investigación (project PID2020-118921RJ-100/AEI/10.13039/501100011033). Isabel Moreno-Indias was supported by the "Miguel Servet Type II" program (CPII21/00013) of the ISCIII-Madrid (Spain), co-financed by the FEDER. The authors thanks for its support of the CIBER-IBIMA-Metagenomics platform, and the Genomics ECAI from IBIMA-Plataforma Bionand.”

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Reviewers' comments:

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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: Partly

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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: Interesting and well-conducted study.

1. The abstract can contain more findings from the molecular analyses (abstracts are different in the system and manuscript).

2. Please interpret more on the bacterial diversity using the Shannon Index.

3. Can include discussion on the potential of Wolbachia as a vector/disease control tool for WNV transmission.

4. Make the rarefaction curve self-explanatory.

Reviewer #2: This research represents the first investigation into the composition of the Cx. Perexiguus microbiota, marking a valuable contribution to the field. However, the findings would be further enriched by the availability of samples collected from multiple locations and at various times.

The results indicate no discernible difference in microbiota composition between WNV-infected and non-infected mosquitoes, nor in their Wolbachia infection rates. Interestingly, the Wolbachia infection rate in Cx. Perexiguus is notably low, potentially increasing susceptibility to WNV infection. The interaction between WNV and Wolbachia infections, however, remains unclear and warrants further exploration.

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Reviewer #1: No

Reviewer #2: No

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Microbiota composition of Culex perexiguus mosquitoes during the West Nile virus outbreak in southern Spain

PONE-D-24-36152R1

Dear Dr. Garrigós,

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.

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Kind regards,

Kelli L. Barr, Ph.D.

Academic Editor

PLOS ONE

Additional Editor Comments (optional): While reviewer 2 was unable to address your revisions, I feel that their comments were addressed sufficiently in the revisions.

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

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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

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

Reviewer #1: 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

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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

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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: (No Response)

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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

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