-->PONE-D-26-11202-->-->Combination of different intervention strategies for Malaria Mosquito vector control in Uganda; a review of secondary data of two districts with moderate to high disease transmission.-->-->PLOS One

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Additional Editor Comments:

The article was reviewed by 2 reviewers and both raised substantial issues requiring major revision

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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: 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: The study is about a very relevant matter, namely, the effectiveness of two major treatments for malaria prevention. It shows interesting findings with sound statistical analysis tools and relevant comparison to similar studies. The conclusions are imporant and merit publication. On the down side, some of the statements made during the paper require further clarification or support. Fore example, in the abstract, it reads "38.5% of reduction rates attributable to variation in either time or deployment of control interventions". It is weird to put time and the type of treatment in the same category. We want to know the percent that is atributable to the control interventions. In the Data Generation section, it seems that only patients that attended medical facilities were measured. This opens the door to the possibility that people test more simply because they have more access to the facilities or clinics. This possible self-selection bias is worth discussing; in a future study, it would be good to try to prevent it by having the data collectors go to places and take samples there, without waiting for the sample to come to them. After graph 2a, it says disapprove, where probably it meant to say disprove. Some comparisons and measures are expressed in ambiguous ways. For example, after Fig 7, it says "the intervention (Acetellic 300

IRS+Royal guard ITNS) is associated with a 16.6 percentage point reduction in malaria

test positivity rates in Yumbe district compared Gulu City". Shall we understand that the intervention was done in both Yumbe and Gulu and the decrease in Yumbe was 16.6 greater than in Gulu? It would be nice to provide confidence intervals for the reduction in each city before and after the intervention.

Other examples of statements that require clarification are the following:

1) "Our findings disagree from a WHO report which

found Clinical trials and pilot studies on dual-insecticide nets to improve malaria control

by 20–50% compared with standard pyrethroid-only nets." With what do your findings disagree exactly? With then umber 20-50%?

2) " Our findings further disagrees

with the additional WHO report that clinical trials in the United Republic of Tanzania

and Benin demonstrated that the pyrethroid-chlorfenapyr nets significantly reduced

malaria infections in children between the ages of 6 months and 10 years." Again, with what part exactly does your study disagree and why?

Reviewer #2: The Authors assessed how different IRS insecticide formulations and ITN types, used alone or in combination, influence malaria test positivity rates in two high-burden districts in Uganda. Studies on the effectiveness of ITNs and/or IRS remain relevant to guide malaria control decisions. However, the manuscript needs to be revised to strengthen the analysis and improve the presentation, interpretation, and discussion of the results.

Major Comments

Given that mosquito ecology and malaria transmission dynamics are strongly influenced by key environmental factors such as rainfall, temperature, humidity, and vegetation indices, I find it concerning that the authors did not account for any climatic factors in their models, particularly in the interrupted time series analysis.

The justification that these data were not readily available is not entirely accurate. Climatic and environmental datasets are widely accessible through remote sensing platforms and could be incorporated into the analysis. For example, temperature and rainfall data can be obtained from sources such as the TerraClimate and CHIRPS, respectively, while vegetation indices like NDVI are available from the Landsat Surface Reflectance-derived-spectral-indices website.

Failing to account for these time-varying factors introduces a high risk of confounding and makes it difficult to attribute observed changes to the interventions. This also limits the validity of the conclusions.

Here is a summary of some additional issues:

#1.The authors consider inter-ITN campaign periods as “no intervention.” This is incorrect because WHO-prequalified ITNs typically remain effective for about 3 years, except in specific cases such as PBO ITNs (with reported durability that averages about 18-24 months in some studies), and their protective effect is expected to last throughout the nets' lifetime. Therefore, observations during these periods should not be classified as no intervention, contrary to the current approach. It is also not clear how the authors incorporated interventions or accounted for intervention coverage in their models.

#2. The parallel trends assumption appears uncertain (particularly in Fig 8). A formal pre-trend test (e.g. treatment × time interaction restricted to the pre-intervention period) should be conducted and reported.

#3. The discussions need revision to strengthen the paper. In this version, the authors mainly compare their findings with other studies but offer limited interpretation and minimal discussion of the implications. From the current analysis and results presented it is clear that the effectiveness of interventions is heavily influenced by the insecticide class and formulation or ITN type. I feel this is an important point that should be emphasised.

#4. One of the main findings is that IRS with Fludora fusion contributed to declines in malaria cases in this study area. This reported impact of Fludora Fusion contrasts with findings from other settings in Uganda (https://doi.org/10.1371/journal.pgph.0003254; https://doi.org/10.1371/journal.pgph.0000676 and https://doi.org/10.21203/rs.3.rs-7518583/v1) where clothianidin-based IRS has shown limited impact. I think this is a significant find, assuming the results do not change after the recommended revisions/analysis and should be discussed in depth. The authors should (i) situate findings within the broader literature, ii) explore possible explanations (e.g. prior IRS exposure, ecological differences) iii) report and interpret the magnitude of effect more explicitly.

#5. It is unclear whether Gulu refers to an entire district or a single city. The authors have used city and district interchangeably. This creates some confusion and raises concerns about comparability. We cannot compare data from a district and that of a township/city.

Minor comments:

#6. The Results section includes elements of study description and discussion. These should be moved to the appropriate sections to maintain a clear and logical structure.

#7. A few observations on terminology, consistency, and tone

- Terminology should be standardised throughout the manuscript. The authors should consistently use either active ingredients, product formulations, or class-level terms, rather than mixing these. For example, alternating between Actellic 300CS and pirimiphos-methyl, or SumiShield/Fludora and clothianidin/deltamethrin, creates confusion. Similarly, there is inconsistency in the level of classification used. In some sections, chemical classes (e.g. “pyrethroid”) are presented alongside specific active ingredients (e.g. “pyriproxyfen”). This should be standardised to either class-level or active-ingredient-level terminology for clarity and consistency

- Authors should consider using either “ITNs” (recommended) or “LLINs” consistently throughout. And should also avoid repeated definitions of IRS and ITNs once introduced, and ensure abbreviations are used consistently after definition.

- Replace words phrasing like “mass scale IRS applications” with standard terminology, either as IRS campaigns or mass ITN distribution

#8. Figures

- Figure 1: Authors should consider using distinct colour shading especially for Gulu and Yumbe.

- Figure 2A:

1. Use clearer distinctions between intervention periods (e.g. colour and line type).

2. Define “M1” and “M7” in a footnote and also add footnotes indicating timing of ITN distributions.

- Figure 8 looks similar to Figure 7. It could be helpful for authors to clarify how they differ or to consider consolidating them if they are redundant.

#9. Tables

- Table 4: revise headings to “Lower 95% CI” and “Upper 95% CI”.

- It would be helpful to Standardise decimal places across all tables and text.

Kindly see the manuscript for additional comments.

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Reviewer #1: No

Reviewer #2: No

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Attachments

Attachment

Submitted filename: Reviewer Comments on Manuscript Number PONE-D-26-11202.pdf

Attachment

Submitted filename: PONE-D-26-11202_Review Feedback.pdf

Combination of different intervention strategies for Malaria Mosquito vector control in Uganda; a review of secondary data of two districts with moderate to high disease transmission.

PONE-D-26-11202R1

Dear Dr. Okek,

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,

Jan Rychtář

Academic Editor

PLOS One

Additional Editor Comments (optional):

All comments addressed

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 #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 #2: Yes

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

Reviewer #2: Yes

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-->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 #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 #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 #2: The authors have addressed the main queries raised. This version is a much-improved version. I would however advise that authors do a thorough check for minor typos and consistency of trade/brand names of ITNs (eg. "PermaNets ITN " vs "PermaNet ITN")

I have no additional comments

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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.

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Reviewer #2: No

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