PONE-D-20-34035

Is Biological Larviciding against Malaria a Starting Point for Integrated Multi-Disease Control? – Observations from a Cluster Randomized Trial in rural Burkina Faso.

PLOS ONE

Dear Dr. Dambach,

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We look forward to receiving your revised manuscript.

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Nicholas C. Manoukis

Academic Editor

PLOS ONE

Additional Editor Comments:

Thank you for your submission- this paper represents substantial field monitoring and treatment application effort, and I believe it provides useful information to programs that include larviciding aiming to control mosquito vectors in W Africa. Please address comments by reviewers.

I find the writing to be understandable but awkward in places- please check through again. In addition, I did not see statistical models presented, though there are p-values in the results section- Please add tables with negative binomial regression results- also provide more information on this model (I am confused as to the mention of a "random effect"- I infer this a mixed model? I feel like more details are needed on statistical approach). On a similar theme, Figures 2 and 4: I think these should include error bars or indication of CI unless I badly misunderstand something.

I am looking forward to seeing a 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: Yes

Reviewer #2: Partly

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

Reviewer #1: Yes

Reviewer #2: I Don't Know

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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: Overall, this is a very straightforward manuscript about the effects of larviciding on the abundance of 2 genera of mosquitoes (the larviciding is originally targeted at a third kind of mosquito in many areas, including this study area). Interestingly, the number of mosquitoes increased from the control year to the treatment year at control sites, and this was reflected in the Culex data as well (further indicating the lack of effect on this genus) - according to Figure 1. I am trying to see how Figure 1 and Figure 2 line up - so it might be easier on the reader to make the colors match between the figures (and even Figure 4).

There are a few copyedit suggestions, check for correct use of hyphens in certain locations (e.g., lines 52, 54, 85) and adding commas in other locations (e.g., lines 56, 83) - not all mentioned here.

I recommend with minor revision - doing some strong copyediting and possibly making the colors of the graphs more uniform so that the reader can more easily connect the results across the graphs.

Reviewer #2: The manuscript requires major revisions particularly in the English composition, cohesion and inclusion of relevant details and references (see comments below). The acceptance of this paper is conditional. The study was done in 2013 and 2014 and only now the authors have submitted this manuscript. Although, WHO still recommends the use of Vectobac WG (AM65-52 strain) as a mosquito larvicide, the authors must show the gap of pertinent studies in the recent past to justify the novelty of their findings.

The manuscript is partly fine, however, it is poorly written and inconclusive. They concluded that:

“Future larviciding programs should be evaluated for including the treatment of Aedes and Culex breeding habitats. Since the major cost components of such programs are labor and transport, other disease vectors could be targeted at little additional cost”.

1) However, their data showed that Baccillus thurigiensis israelensis (Bti) larviciding had limited or no impact against Culex mosquitoes. Their major finding was Bti larviciding reduced up to 34% of Aedes mosquitoes in breeding sites in public spaces.

2) Based on their indoor and outdoor captures of adult mosquitoes, they found that Anopheles were reduced at indoor captures twice as high compared to those from outdoors (lines 224-226). But they were not certain what factors that led to higher reduction of Culex quinquefasciatus and Aedes aegypti at outdoors (lines 224-228).

The above major findings might be the reason that the title poses a question implying that the results are not conclusive and their underlying purpose of this submission apparently is to report their observations of their first year of intervention year. Hence, they “concluded” with a recommendation to further evaluate future larviciding programs for including the treatment of Aedes and Culex breeding sites with the use of Bti larvicide.

Note that the scope of this manuscript included only the baseline and the first intervention year. The paper might be more conclusive if the results of the second intervention year would be included to be more certain on their conclusion.

I suggest that a statistician’s opinion can further elucidate the judgment of their statistical analyses. My comments include:

1. Lines 169-171: Please describe clearly and explicitly the interpretations of the statistical analyses performed. What were the Culex mosquitoes significant of? How about Aedes and Mansonia?

2. Lines 257-263: The sample size bias of 122 Aedes and 150 Culex mosquitoes was justified by stating Harbach (2012). I am not certain whether their justification is statistically acceptable. What I am certain is in their research design, the power analysis of sample size was not predetermined prior to actual field work, hence, they had to retrospectively justify what they got during the actual field work.

Add additional relevant references that can augment the use of Bti in setting the background of your study. Were there related studies in the recent past that used Bti against malaria mosquitoes in other African countries and in Asia? How novel is your work particularly that your study was done in 2013 and 2014 yet? Lines 46-49: Reference 1 is published in 2007 and is not current anymore, inconsistent of the sentence in these lines.

Lines 52, 56, 61-62, 75, 210-214, 219, etc.: Improve grammar on the agreement between subject and verb, and sentence construction.

Methods:

1. Please include a GIS-map of the study site. Indicate the months of the two seasons in the study sites.

2. Line 98, 239, 245-246, 252: References please. Lines 267-268: Please cite relevant references indicating the average flight distances of each mosquito species included in the study. Avoid generalizing them.

3. Lines 99-101: Please describe briefly but concisely the third larviciding choice (risk map-based larvicide). What do you mean by the most productive breeding sites? Please cite references.

4. Line 142: Please indicate the primers and PCR-reaction mixture used in identifying the sibling species of Culex even though you cited Kasai (2008) and Smith-Fonseca (2004).

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Reviewer #1: No

Reviewer #2: No

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Is Biological Larviciding against Malaria a Starting Point for Integrated Multi-Disease Control? – Observations from a Cluster Randomized Trial in rural Burkina Faso.

PONE-D-20-34035R1

Dear Dr. Dambach,

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,

Nicholas C. Manoukis

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

Congratulations on a very nice revision. The paper is, in my view, acceptable for publication in PLOS ONE. The funding statement should be fine in the Acknowledgments, unless editorial staff have other information.

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

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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 #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: My previous comments regarding readability / English language and grammar have been well-addressed. The figures have also been improved, with a more consistent color scheme (if still not perfectly harmonized across all figures). The statistical questions raised by the other reviewer appear to have been addressed as well, and additional references/citations included.

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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