Larvicidal properties of terpenoid-based nanoemulsions against the dengue vector Aedes aegypti L. and their potential toxicity against non-target organism

Jonatas Lobato Duarte; Stéphane Duchon; Leonardo Delello Di Filippo; Marlus Chorilli; Vincent Corbel

doi:10.1371/journal.pone.0293124

Peer Review History

Original SubmissionJune 3, 2023
26 Jun 2023 Decision Letter - Mozaniel Santana de Oliveira, Editor PONE-D-23-16634Larvicidal properties of terpenoid-based nanoemulsions against the Dengue vector Aedes aegypti L. and their potential toxicity against non-target organismPLOS ONE Dear Dr. Corbel, 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. Please submit your revised manuscript by Aug 10 2023 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file. 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The PLOS ONE style templates can be found at https://journals.plos.org/plosone/s/file?id=wjVg/PLOSOne_formatting_sample_main_body.pdf and https://journals.plos.org/plosone/s/file?id=ba62/PLOSOne_formatting_sample_title_authors_affiliations.pdf 2. Thank you for stating the following financial disclosure: "This project has received funding from Sao Paulo research foundation-FAPESP under the grants n°2021/11487-4 and 2019/25125-7. This project also receives a co-funding from the European Union HORIZON-MSCA-2021-SE-01 (INOVEC project), under the grant n°101086257. " Please state what role the funders took in the study. If the funders had no role, please state: "The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript." If this statement is not correct you must amend it as needed. Please include this amended Role of Funder statement in your cover letter; we will change the online submission form on your behalf. 3. Please expand the acronym “FAPESP” and "MSCA" (as indicated in your financial disclosure) so that it states the name of your funders in full. This information should be included in your cover letter; we will change the online submission form on your behalf. 4. We note that you have stated that you will provide repository information for your data at acceptance. Should your manuscript be accepted for publication, we will hold it until you provide the relevant accession numbers or DOIs necessary to access your data. If you wish to make changes to your Data Availability statement, please describe these changes in your cover letter and we will update your Data Availability statement to reflect the information you provide. [Note: HTML markup is below. Please do not edit.] 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 ******** 2. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: Yes Reviewer #2: No ****** 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: No ****** 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 ****** 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: Good manuscript and well written. The concept is good as alternative method in insect control technology. The study has detailed view and experiments in terms of proving their concepts. Please refer to my comments for further improvement of this manuscripts. Reviewer #2: The manuscript by Duarte et al. brings a straightforward presentation of the effects of two terpenoids and their nanoemulsions against Aedes aegypti larvae, and the test against a non-target organism. It is a well conducted and objective study. However, the presentation of data is very poor. I included some comments in the PDF file, but basically the main problems are: 1. Legends of Tables and figures are almost non-existant. There is no description of the meaning of variables, measurements, statistical tests and symbols, experimental designs. Please provide complete Legends. 2. Statistical analysis was performed without adequate report of results. Fos example, in some cases ANOVA was performed without reference to normality of data sets, which I think it is hard to demonstrate with N=5. Please provide proper statistical analysis. ****** 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. 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: Yes: Rajiv Ravi (PhD), Sumitomo Chemical Enviro-Agro Pacific Sdn.Bhd Reviewer #2: No ******** [NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.] While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/. PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at figures@plos.org. Please note that Supporting Information files do not need this step. Attachments Attachment Submitted filename: Comments Reviewer.docx Attachment Submitted filename: PONE-D-23-16634_rev.pdf https://doi.org/10.1371/journal.pone.0293124.r001
Revision 1
7 Jul 2023 Author Response We would like to express our sincere gratitude for the valuable feedback provided by the academic editor and reviewer(s) regarding our manuscript entitled " Larvicidal properties of terpenoid-based nanoemulsions against the Dengue vector Aedes aegypti L. and their potential toxicity against non-target organism." We appreciate their time and effort in thoroughly assessing our work. In response to their insightful comments, we have carefully considered each point raised and offer our rebuttal and revisions as follows: REVIEWER #1 Something seems wrong here. Why the total is not 100%? A: We corrected the information in the main text. What was the light exposure? A: We used a light/dark cycle of 12 hours. We added this information in the methods section. What is the meaning of AHI? A: We corrected the translation mistake in the main text. The correct acronym is IHC. One way ANOVA with N=5 do not seem reasonable. how the authors tested the normality of distribution? A: We used the Shapiro wilk test. Decide between BORA or Bora and correct it along the manuscript. A: Thank you for the suggestion. We standardized “Bora” throughout the text. Where is this data? refer it in the text please. A: We included a reference to this data in the text. The legend of this Table is surprisingly poor. It´s not possible to know what p (italics and lowercase, please) means, or which statistical test was performed, or even the number of biological replicas (n is number of mosquitoes?). Please provide explanation for this data and a better statistical report (use addiditonal tables if necessary). A: We reformulated the legend, adding the necessary information for data interpretation. Which test was used for the normality of distribution? ANOVA implies this condition. A: We used the Shapiro wilk test to check the normality of the data. Why GC? A: We corrected the translation mistake in the main text. Other strategies of stabilization of essential oils by encapsulation were alerady proposed, for example yeast encapsulation, as demonstrated in: 1. Parasit Vectors. 2021 May 22;14(1):272. doi: 10.1186/s13071-021-04733-2. 2. Parasit Vectors. 2020 Jan 13;13(1):19. doi: 10.1186/s13071-019-3870-4. A: We appreciate the suggestion. We would like to reiterate that the objective of this article is to discuss approaches related to nanoemulsions and larvicidal activity. At this time, we have chosen not to include other approaches in the manuscript. REVIEWER #2 Introduction Good explanations on the introduction about the mechanisms of resistant and objective of this study. Very clear and on the points for a research statements. The experiment design is much targeted and design well. Appropriate in this context. Line 56-65 can be mentioned more comprehensive, especially the alternative method. You are not using those method and not relevant in your study scope A: Thank you for the suggestion. We have chosen to remove that section from the manuscript. Explain more on the Nano emulsions technology for slow release. How the mechanisms work for slow release for a highly volatile component. Line 92-96 A: We appreciate the reviewer's comment. We have included a detailed explanation on Nano emulsion technology for slow release in the manuscript. Materials and Method If you have pictures of behavioral analysis and mortality of fish, please include. This will be a good reference for repeating this kind of experiment. A: Thank you for your suggestion. While we understand the value of including pictures of behavioral analysis and fish mortality in the manuscript, we would like to clarify that these specific visuals are not available in our study. However, we have provided detailed descriptions of the behavioral analysis and fish mortality results in the text. Additionally, we have referenced relevant literature that provides visual documentation of similar experiments, which can serve as a valuable reference for researchers interested in replicating these experiments. Explain the line 186-193 with the picture for each Stage I-III A:Thank you for your suggestion. While we understand the value of including pictures of behavioral analysis and fish mortality in the manuscript, we would like to clarify that these specific visuals are not available in our study. Results Table 2 and Table 3, please include in title the RR ratio index range based on WHO reference. Example; According to WHO RR < 5 indicates a susceptible field population; RR from 5-10 indicates moderate resistance and RR > 10 indicates high resistance, (Monitoring and managing insecticide resistance in Aedes mosquito populations. Interim guidance for entomologists. WHO/ZIKV/VC/16.1. Geneva: World Health Organization; 2016) A: We included this information in the method section as this is not a result (P148-150). We hope that these revisions adequately address the concerns raised and align the manuscript more closely with the objectives and scope of PLOS ONE. We are confident that the revised version now meets the high standards of academic excellence set by your esteemed journal. Once again, we would like to express our sincere appreciation for the thorough review and constructive feedback provided by the academic editor and reviewer(s). Their expertise and guidance have undoubtedly improved the quality of our work. Thank you for considering our revised manuscript. We look forward to receiving your final decision regarding its publication. Yours sincerely, Vincent Corbel Attachments Attachment Submitted filename: Response to Reviewers.docx https://doi.org/10.1371/journal.pone.0293124.r002
31 Jul 2023 Decision Letter - Mozaniel Santana de Oliveira, Editor PONE-D-23-16634R1 Larvicidal properties of terpenoid-based nanoemulsions against the Dengue vector Aedes aegypti L. and their potential toxicity against non-target organism PLOS ONE Dear Dr. Corbel, Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we have decided that your manuscript does not meet our criteria for publication and must therefore be rejected. I am sorry that we cannot be more positive on this occasion, but hope that you appreciate the reasons for this decision. Kind regards, Mozaniel Santana de Oliveira, Ph.D Academic Editor PLOS ONE [Note: HTML markup is below. Please do not edit.] 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 #3: (No Response) ******** 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 #3: (No Response) ****** 3. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: Yes Reviewer #3: (No Response) ****** 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 #3: (No Response) ****** 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 #3: (No Response) ****** 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: 1. All the questions has been answered and acceptable for publications 2. Format is acceptable 3. Manuscript is acceptable Reviewer #3: The authors did not make the necessary changes for the manuscript to be published in this journal. ****** 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: Yes: Rajiv Ravi Reviewer #3: No ******** [NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.] - - - - - For journal use only: PONEDEC3 Attachments Attachment Submitted filename: REVIEWER2.docx https://doi.org/10.1371/journal.pone.0293124.r003
Revision 2
17 Aug 2023 Author Response We would like to express our sincere gratitude for the valuable feedback provided by the academic editor and reviewer(s) regarding our manuscript entitled " Larvicidal properties of terpenoid-based nanoemulsions against the Dengue vector Aedes aegypti L. and their potential toxicity against non-target organism." We appreciate their time and effort in thoroughly assessing our work. In response to their insightful comments, we have carefully considered each point raised and offer our rebuttal and revisions as follows: REVIEWER #1 Something seems wrong here. Why the total is not 100%? A: We corrected the information in the main text. What was the light exposure? A: We used a light/dark cycle of 12 hours. We added this information in the methods section. What is the meaning of AHI? A: The correct acronym is IHC (Index of Histopathological Changes). We corrected the mistake in the text. One way ANOVA with N=5 do not seem reasonable. how the authors tested the normality of distribution? A: We used the Shapiro wilk test to check the distribution. The data showed normal distribution (Statistic 0.9598, P value: 0.6577). Decide between BORA or Bora and correct it along the manuscript. A: Thank you for the suggestion. We standardized “Bora” throughout the text. Where is this data? refer it in the text please. A: We included a reference to this data in the text. The legend of this Table is surprisingly poor. It´s not possible to know what p (italics and lowercase, please) means, or which statistical test was performed, or even the number of biological replicas (n is number of mosquitoes?). Please provide explanation for this data and a better statistical report (use addiditonal tables if necessary). A: We reformulated the legend, adding the necessary information for data interpretation. Which test was used for the normality of distribution? ANOVA implies this condition. A: We used the Shapiro wilk test to check the normality of the data. The data showed normal distribution (Statistic 0.9598, P value: 0.6577). Why GC? A: We corrected the translation mistake in the main text. Other strategies of stabilization of essential oils by encapsulation were already proposed, for example yeast encapsulation, as demonstrated in: 1. Parasit Vectors. 2021 May 22;14(1):272. doi: 10.1186/s13071-021-04733-2. 2. Parasit Vectors. 2020 Jan 13;13(1):19. doi: 10.1186/s13071-019-3870-4. A: We appreciate the suggestion. We inserted the suggested sentence and references in the manuscript. REVIEWER #2 Introduction Good explanations on the introduction about the mechanisms of resistant and objective of this study. Very clear and on the points for research statements. The experiment design is much targeted and design well. Appropriate in this context. Line 56-65 can be mentioned more comprehensive, especially the alternative method. You are not using those method and not relevant in your study scope A: Thank you for the suggestion. We have chosen to remove that section from the manuscript. Explain more on the Nano emulsions technology for slow release. How the mechanisms work for slow release for a highly volatile component. Line 92-96 A: We appreciate the reviewer's comment. We have included a detailed explanation on Nano emulsion technology for slow release in the manuscript (see introduction). Materials and Method If you have pictures of behavioral analysis and mortality of fish, please include. This will be a good reference for repeating this kind of experiment. A: Thank you for your suggestion. While we understand the value of including pictures of behavioral analysis and fish mortality in the manuscript, we would like to clarify that these specific visuals are not available in our study. However, we have provided detailed descriptions of the behavioral analysis and fish mortality results in the text. Additionally, we have referenced relevant literature that provides visual documentation of similar experiments, which can serve as a valuable reference for researchers interested in replicating these experiments. Explain the line 186-193 with the picture for each Stage I-III A:Thank you for your suggestion. While we understand the value of including pictures of behavioral analysis and fish mortality in the manuscript, we would like to clarify that these specific visuals are not available in our study. Results Table 2 and Table 3, please include in title the RR ratio index range based on WHO reference. Example; According to WHO RR < 5 indicates a susceptible field population; RR from 5-10 indicates moderate resistance and RR > 10 indicates high resistance, (Monitoring and managing insecticide resistance in Aedes mosquito populations. Interim guidance for entomologists. WHO/ZIKV/VC/16.1. Geneva: World Health Organization; 2016) A: We included this information in the method section as this is not a result (P148-150). We hope that these revisions adequately address the concerns raised and align the manuscript more closely with the objectives and scope of PLOS ONE. We are confident that the revised version now meets the high standards of academic excellence set by your esteemed journal. Once again, we would like to express our sincere appreciation for the thorough review and constructive feedback provided by the academic editor and reviewer(s). Their expertise and guidance have undoubtedly improved the quality of our work. Thank you for considering our revised manuscript. We look forward to receiving your final decision regarding its publication. Yours sincerely, Vincent Corbel Attachments Attachment Submitted filename: Response to Reviewers 1 and 2.docx https://doi.org/10.1371/journal.pone.0293124.r004
6 Oct 2023 Decision Letter - Pedro L. Oliveira, Editor Larvicidal properties of terpenoid-based nanoemulsions against the Dengue vector Aedes aegypti L. and their potential toxicity against non-target organism PONE-D-23-16634R2 Dear Dr. Corbel, 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. An invoice for payment will follow shortly after the formal acceptance. To ensure an efficient process, please log into Editorial Manager at http://www.editorialmanager.com/pone/, click the 'Update My Information' link at the top of the page, and double check that your user information is up-to-date. If you have any billing related questions, please contact our Author Billing department directly at authorbilling@plos.org. If your institution or institutions have a press office, please notify them about your upcoming paper to help maximize its impact. If they’ll be preparing press materials, please inform our press team as soon as possible -- no later than 48 hours after receiving the formal acceptance. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information, please contact onepress@plos.org. Kind regards, Pedro L. Oliveira Academic Editor PLOS ONE Additional Editor Comments (optional): Reviewers' comments: https://doi.org/10.1371/journal.pone.0293124.r005
Formally Accepted
11 Oct 2023 Acceptance Letter - Pedro L. Oliveira, Editor PONE-D-23-16634R2 Larvicidal properties of terpenoid-based nanoemulsions against the Dengue vector Aedes aegypti L. and their potential toxicity against non-target organism Dear Dr. Corbel: I'm pleased to inform you that your manuscript has been deemed suitable for publication in PLOS ONE. Congratulations! Your manuscript is now with our production department. If your institution or institutions have a press office, please let them know about your upcoming paper now to help maximize its impact. If they'll be preparing press materials, please inform our press team within the next 48 hours. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information please contact onepress@plos.org. If we can help with anything else, please email us at plosone@plos.org. Thank you for submitting your work to PLOS ONE and supporting open access. Kind regards, PLOS ONE Editorial Office Staff on behalf of Dr. Pedro L. Oliveira Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0293124.r006

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