-->PONE-D-25-50468-->-->Balancing pest control and natural enemy conservation: Seed Treatment Effects in Cotton-->-->PLOS One

Dear Dr. Yalçın,

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.

-->-->When revising your manuscript and as requested by reviewer 2, please give special consideration to (Abstract, supporting materials, double check figures/tables references and discussion flow). Also you should well describe in detail the sweeping methods (trapped insects and stages) in Mat&Met section.-->--> -->-->Good luck-->-->

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Rachid Bouharroud

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: Partly

Reviewer #2: Yes

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

Reviewer #1: I Don't Know

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.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.-->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.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?

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Reviewer #1: No

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: Introduction is fairly long. A number of studies are cited, indicating a lot of literature already available on this topic. What are objectives of this study needs to be enlisted and introduction should be shaped according to them to build argument by identifying gaps rather than reviewing earlier literature.

L102: For the experimental design, three replicates were run following a randomized complete block design. In a small-scale study of this kind, different field as replicates may be more appropriate to include variation as true replicates.

L119: Sweep nests were used to assess abundance. If this method was used against flying adults or also when larvae or immobile or less immobile stages were present, needs clarity on this. how much time was spent per sweep.

Results: Seasonal counts can be compared among treatments at the end of season. Line graphs can be made supplementary and give their seasonal means comparison, results are too long and hard to follow under current data representation. Also provide information on dominant species of pests and predators.

Discussion: What authors find new in this study and discuss the way forward. What are the limitation of this study and possible research direction?

Reviewer #2: This two-year field study provides a valuable community-level assessment of the impact of two common seed treatments—clothianidin (CLO, insecticide) and azoxystrobin + metalaxyl-m + fludioxonil (AMF, fungicide combination)—on arthropod pests and their natural enemies in a cotton agroecosystem. The research addresses an important gap by moving beyond single-species assessments to evaluate whole-community dynamics, which is crucial for developing sustainable Integrated Pest Management (IPM) strategies.

Key Strengths of the Manuscript:

Strong Experimental Design: A two-year, replicated field trial with an untreated control provides robust, ecologically relevant data.

Comprehensive Community Analysis: The use of Principal Response Curve (PRC) analysis to synthesize community-level effects is a significant methodological strength, clearly visualizing how entire arthropod assemblages shift in response to treatments over time.

Important Findings: The study convincingly demonstrates that both CLO and AMF seed treatments significantly reduce the abundance of key natural enemy families (e.g., Nabidae, Chrysopidae, Coccinellidae) while also suppressing pest populations like Cicadellidae. The documented negative correlation between pest and predator suppression highlights a critical trade-off in pest management.

Clear Conclusions: The conclusions are well-supported by the data, emphasizing that seed treatments are not "safe" for natural enemies and calling for more judicious, need-based use within IPM frameworks.

Suggestions for Revision and Clarification:

Title Refinement: The current title ("Balancing pest control and natural enemy conservation: Seed Treatment Effects in Cotton") is slightly generic. Consider making it more specific to your key finding, e.g., "Trade-offs in cotton pest management: Seed treatments suppress pests but reduce natural enemy abundance in the arthropod community."

Abstract Enhancement: The abstract should more clearly state the main takeaway. Explicitly mention that your study found a significant reduction in predator abundance alongside pest suppression, creating a potential trade-off. Briefly specify the two treatments used.

Results Presentation: The text frequently refers to figures and tables (e.g., Fig 1, S1 Table) that are not included in the provided text file. Ensure all in-text citations have corresponding, clearly labeled items in the submitted manuscript.

Discussion Flow: The discussion is comprehensive but could be more tightly structured. Consider subheadings (e.g., "Effects on Pest Communities," "Effects on Predator Communities," "Community-Level Implications") to improve readability and guide the reader through your interpretations.

Clarity on "Neonicotinoid" Terminology: Clothianidin is a neonicotinoid. The introduction and discussion mention neonicotinoids broadly but could more explicitly frame CLO within this well-known class from the outset, as their non-target effects are a major point of discussion.

Statistical Reporting Consistency: Ensure that all statistical results (F-values, degrees of freedom, p-values) are reported consistently (e.g., F(1, 2) = 5.90, p = 0.038).

Data Availability Statement: The statement "The data supporting this study's findings are available in this article's supplementary material" is good, but PLOS ONE requires that the data be fully available without restriction. Double-check that all raw data underlying the figures and analyses are indeed included in the submitted Supporting Information files.

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Reviewer #1: No

Reviewer #2: No

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Trade-offs in cotton pest management: Seed treatments suppress pests but reduce the abundance of natural enemies in the arthropod community.

PONE-D-25-50468R1

Dear Dr. Yalçın,

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,

Rachid Bouharroud

Academic Editor

PLOS One

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