Field and laboratory assessment of larvicidal activity of tobacco plants and the cigarette butt waste on Culex pipiens (Linnaeus, 1758), Aedes aegypti (Linnaeus, 1762) L. and non-target organisms

Mohamed M. Baz; Yasser A. El-Sayed; Samah Abdel Gawad; Mohammed E. Gad; Moustafa Ibrahim; Mohammed H. Alruhaili; Hattan S. Gattan; Reham M. Mostafa; Sarita Kumar; Ibrahim E. Hussein; Abdelfattah Selim; Abeer M. Alkhaibari; Ahlam F. Moharam

doi:10.1371/journal.pone.0333549

Peer Review History

Original SubmissionJune 23, 2025
5 Aug 2025 Decision Letter - Jorddy Neves Cruz, Editor PONE-D-25-33977Tobacco plant and cigarette butt wastes: field and laboratory evaluation of insecticidal activities against Culex pipiens, Aedes aegypti (Diptera: Culicidae), and non-target organismsPLOS ONE Dear Dr. Selim, 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. ============================== ACADEMIC EDITOR NOTES: Dear authors, all the referees raised important questions about the manuscript. I suggest that the authors conduct a thorough review of the manuscript; the response letter should contain a point-by-point answer to all the referees' questions. Furthermore, all changes to the text should be highlighted in yellow. ============================== Please submit your revised manuscript by Sep 19 2025 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. Please include the following items when submitting your revised manuscript: A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'. A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'. An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'. If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter. If applicable, we recommend that you deposit your laboratory protocols in protocols.io to enhance the reproducibility of your results. Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: https://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols. Additionally, PLOS ONE offers an option for publishing peer-reviewed Lab Protocol articles, which describe protocols hosted on protocols.io. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols. We look forward to receiving your revised manuscript. Kind regards, Jorddy Neves Cruz Academic Editor PLOS ONE Journal requirements: When submitting your revision, we need you to address these additional requirements. 1. Please ensure that your manuscript meets PLOS ONE's style requirements, including those for file naming. 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. In your Methods section, please provide additional information regarding the permits you obtained for the work. Please ensure you have included the full name of the authority that approved the field site access and, if no permits were required, a brief statement explaining why. 3. In the online submission form, you indicated that [data available on request form corresponding author]. All PLOS journals now require all data underlying the findings described in their manuscript to be freely available to other researchers, either 1. In a public repository, 2. Within the manuscript itself, or 3. Uploaded as supplementary information. This policy applies to all data except where public deposition would breach compliance with the protocol approved by your research ethics board. If your data cannot be made publicly available for ethical or legal reasons (e.g., public availability would compromise patient privacy), please explain your reasons on resubmission and your exemption request will be escalated for approval. 4. If the reviewer comments include a recommendation to cite specific previously published works, please review and evaluate these publications to determine whether they are relevant and should be cited. There is no requirement to cite these works unless the editor has indicated otherwise. Additional Editor Comments (if provided): [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: No Reviewer #2: Yes Reviewer #3: Yes ******** 2. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: Yes Reviewer #2: Yes Reviewer #3: Yes ****** 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 Reviewer #3: Yes ****** 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: No Reviewer #2: Yes Reviewer #3: 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: Comments to the authors The manuscript titled “Tobacco plant and cigarette butt wastes: field and laboratory evaluation of insecticidal activities against Culex pipiens, Aedes aegypti (Diptera: Culicidae), and non-target organisms”. The present study evaluated about to show that certain chemicals in the tobacco plant (Nicotiana 34 tabacum) and cigarette butt wastes (CBWs) are crucial in biological processes against mosquitoes. After carefully reviewing this manuscript I can not see merit in this research and also the current form contains several writing, scientific, grammatical and typographical errors so please carefully correct the errors based on the following comments; 1. The manuscript title is hard to follow, rewrite the title. 2. Line 26-33: scientifically not sound. 3. Line 33-37:the objective and methodology not clear. 4. The end of the abstract the conclusion is missing. 5. If possible add a few more keywords and avoid the keyword from the manuscript title. 6. Line 51-83: scientifically not sound, the current form looks like a textbook. Please carefully revise it. 7. Line 87: references are missing. 8. Line 91: add a few more recent references. 9. Line 91-92: hard to follow. 10. The entire manuscript is scientifically not sound and hard to follow. 11. In the introduction the rationale of present study is not properly described. 12. Provide latitude and longitude information of plant sample collected areas. 13. The plant sample authenticated by any plant taxonomist? If yes, provide the details. If not then how did you identify the plant? 14. Line 132-133: provide the room temperature and humidity details. 15. Line 137: why methanol solvent? What about other solvents? 16. Did you extract the Cigarette butt? Or you extracted Cigarette? If you extracted Cigarettes, how do you recycle the Cigarette butt? I think the author is confused with this research. 17. Line 196: what is the nutrient content of Tetramin? 18. Line 196-197: not clear. 19. Line 203: typo error. 20. The methodological part is poorly written, the current form of the section scientifically not sound. So revise this part with proper references, I do not see any proffer reference in the methods section. 21. The results part must be improve 22. In the discussion part scientifically not sound and don’t provide your results in the discussion part. This is not the right place to write your results. 23. The reference section contains several formatting and typographical errors so carefully correct the errors. 24. Throughout the manuscript I can see several scientific, technical, fundamental, grammatical and typographical errors so please carefully correct. Reviewer #2: The manuscript entitled “Tobacco plant and cigarette butt wastes: field and laboratory evaluation of insecticidal activities against Culex pipiens, Aedes aegypti (Diptera: Culicidae), and non-target organisms” by Baz et al. is well-written in simple and clear English and presents valuable information. While reading through the manuscript, I found it easy to understand due to the straightforward language used. Below are a few comments from my side: 1. All scientific names must be italicized and given in full at first mention (e.g., Aedes aegypti Linnaeus). Afterwards, abbreviate genus (e.g., Ae. aegypti). In the title, include the author citation (“Say”) after Aedes aegypti. 2. Abbreviations should be expanded upon their first use. 3. Please provide the author citation (taxonomic authority) for the mosquito species in the title (e.g., “Say”). 4. The Introduction is well-written but could be enhanced by including more detailed information about the tobacco plant, including nicotine and other compounds it contains. 5. The Introduction would benefit from more detail on tobacco’s alkaloids (nicotine, nornicotine, anatabine), flavonoids and terpenes, including approximate concentrations in Nicotiana tabacum L. leaves. 6. Correct the repeated name in line 112. 7. Cigarette butts contain many toxic compounds, such as arsenic, a heavy metal. Highlighting the insecticidal potential of cigarette butt-derived products could strengthen the manuscript. "What about the toxicity of the present compounds? Is it reliable and safe to use them as insecticides?". 8. Add a sentence in the Introduction (or early Discussion) succinctly stating how cigarette‑butt–derived larvicides compare in cost, environmental impact, or ease of production versus conventional options. 9. The Materials and Methods and Results sections are well-structured and clearly presented. 10. Was any standard control (Pesticide) used for comparison in this study? Please clarify. 11. Figures 4 to 8 currently show mean values only. Add standard error or confidence interval bars to all bar charts and line plots to indicate variability. 12. Was ethical clearance obtained for the use of hamster mouse in this study? If so, please provide the relevant details in the section 2.3.1. 13. The Discussion section is well-structured but is overly lengthy. Consider condensing or reorganizing it for better flow and add the limitation and future direction of the study. 14. The Conclusion should be rewritten. Please summarize the core findings and include future perspectives. 15. Cross‑check that all figures/tables are cited in order. 16. The entire manuscript should be thoroughly checked for grammatical, typographical, and spacing errors. Reviewer #3: 1. Lines 26 to 33 of the abstract should be rewritten in more concise and informative way. 2. The abstract lacks the main phytochemical results. Authors should highlight the main identified phytoconstituents. 3. In the plant collection part, kindly add the name of the botanist who authenticated the collected tobacco plant. 4. In lines 182 and 187, why only the plant extract and one brand were selected for analysis rather than the other collected cigarette butt samples? 5. Upon collecting the cigarette butts from public areas, how could the authors differentiate the exact brand of each of them? 6. The authors are advised to check for any Grammatik or spelling mistakes throughout the manuscript. 7. Line 352, the authors mistype “acetone extract” although the study is for methanol extract. correct please. 8. A standardization method, using HPLC for example, for the percentage of different phytochemicals present in the cigarette butt extracts is requested. 9. The discussion part is too long and need to be rewritten in more informative way. 10. The discussion part may include the authors’ opinion about the feasibility of using cigarette butt waste as a source for bio insecticide. 11. In the references part, the latin names should be italic. 12. In table 9, instead of bioactive compounds in the header, the authors may use chemical class or class. 13. In fig. 2 kindly add the scale used in capturing this picture. 14. Kindly provide an explanation for figure 10, the FTIR results, in the results section. ****** 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: No Reviewer #2: No 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.] 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. https://doi.org/10.1371/journal.pone.0333549.r001
Revision 1
27 Aug 2025 Author Response Respected Prof. Dr. ………… Thanks to the Editorial board and reviewers for their careful and valuable review, including comments, opinions, directions, and their kind words about the manuscript. • More recent information was added to the text sections. • All manuscript sections were improved as recommended. • Corrections were highlighted in yellow color. We appreciate and value the effort you put into our manuscript to make it satisfactory and high quality enough for publication in your respected journal. Reviewer #1: Comments to the authors The manuscript titled “Tobacco plant and cigarette butt wastes: field and laboratory evaluation of insecticidal activities against Culex pipiens, Aedes aegypti (Diptera: Culicidae), and non-target organisms”. The present study evaluated about to show that certain chemicals in the tobacco plant (Nicotiana 34 tabacum) and cigarette butt wastes (CBWs) are crucial in biological processes against mosquitoes. After carefully reviewing this manuscript I can not see merit in this research and also the current form contains several writing, scientific, grammatical and typographical errors so please carefully correct the errors based on the following comments; Response: Thank you for your careful review and constructive comments. 1. The manuscript title is hard to follow, rewrite the title. Response: Checked and rewritten. 2. Line 26-33: scientifically not sound. Response: Revised and corrected (Page 2, line 27-36). 3. Line 33-37: the objective and methodology not clear. Response: Revised and corrected (Page 2, line 31-36). 4. The end of the abstract the conclusion is missing. Response: Revised and added (Page 2, line 48-49). 5. If possible, add a few more keywords and avoid the keyword from the manuscript title. Response: Revised and added (Page 2, line 50-51). 6. Line 51-83: scientifically not sound, the current form looks like a textbook. Please carefully revise it. Response: Revised and Corrected (Page 3, line 53-70). 7. Line 87: references are missing. Response: Revised and added (Page 4, line 88). 8. Line 91: add a few more recent references. Response: Revised and added (Page 5, line 107). 9. Line 91-92: hard to follow Response: Revised and corrected (Page 4, line 92-94). 10. The entire manuscript is scientifically not sound and hard to follow. Response: Checked and revised. 11. In the introduction the rationale of present study is not properly described. Response: Revised and added (Page 5, line 95-109). 12. Provide latitude and longitude information of plant sample collected areas. Response: Revised and added (Page 60, line 139). 13. The plant sample authenticated by any plant taxonomist? If yes, provide the details. If not then how did you identify the plant? Response: Revised and added (Page 6, line 143-144). 14. Line 132-133: provide the room temperature and humidity details. Response: Revised and added (Page 6, line 144). 15. Line 137: why methanol solvent? What about other solvents? Response: Methanol is used as a solvent for plant extractions because of its high polarity, which enables it to extract a wide range of bioactive plant compounds, such as alkaloids and phenols. It also effectively penetrates plant cell walls, increasing the yield of extracts. In addition to its easy evaporation and compatibility with analytical techniques. Furthermore, it does not require other solvents in our study for the variety of cigarette butts and tobacco plants. So, methanol is a practical and economical choice for laboratory research, as demonstrated in previous research 16. Did you extract the Cigarette butt? Or you extracted Cigarette? If you extracted Cigarettes, how do you recycle the Cigarette butt? I think the author is confused with this research. Response: Sir, there is no confusion, as the remaining tobacco was extracted from the cigarette, represented by the cigarette butt, and this is what was explained in Figure 3. 17. Line 196: what is the nutrient content of Tetramin? Response: Tetramin fish food is a commercial food commonly used in fish and aquatic worm farming. It is rich in proteins, vitamins, and minerals that support the growth of insects and fish, including mosquito larvae in laboratory experiments. Tetramin fish food is a commonly used compound in laboratories that breed mosquitoes for research purposes. 18. Line 196-197: not clear. Response: Revised and corrected. 19. Line 203: typo error. Response: Revised and corrected. 20. The methodological part is poorly written, the current form of the section scientifically not sound. So revise this part with proper references, I do not see any proffer reference in the methods section. Response: Revised and corrected. 21. The results part must be improve Response: Checked and corrected. 22. In the discussion part scientifically not sound and don’t provide your results in the discussion part. This is not the right place to write your results. Response.: Discussion part has been revised as per the suggestions. 23. The reference section contains several formatting and typographical errors so carefully correct the errors. Response: Care has been taken to remove errors. 24. Throughout the manuscript I can see several scientific, technical, fundamental, grammatical and typographical errors so please carefully correct. Response: The manuscript has been carefully revised. Thanks for your suggestions. Reviewer #2: The manuscript entitled “Tobacco plant and cigarette butt wastes: field and laboratory evaluation of insecticidal activities against Culex pipiens, Aedes aegypti (Diptera: Culicidae), and non-target organisms” by Baz et al. is well-written in simple and clear English and presents valuable information. While reading through the manuscript, I found it easy to understand due to the straightforward language used. Below are a few comments from my side: Response: Thank you for your careful review and constructive comments. 1. All scientific names must be italicized and given in full at first mention (e.g., Aedes aegypti Linnaeus). Afterwards, abbreviate genus (e.g., Ae. aegypti). In the title, include the author citation (“Say”) after Aedes aegypti. Response: Revised and corrected. Sir, Carl Linnaeus first described Culex pipiens (Linnaeus) in 1758, while Aedes aegypti (Linnaeus) was described in 1762. Therefore, they are the original and formal scientific name given by Linnaeus. 2. Abbreviations should be expanded upon their first use. Response: Checked and corrected. 3. Please provide the author citation (taxonomic authority) for the mosquito species in the title (e.g., “Say”). Response: Revised and corrected. 4. The Introduction is well-written but could be enhanced by including more detailed information about the tobacco plant, including nicotine and other compounds it contains. Response: Revised and added (Page 4, line 95-109). 5. The Introduction would benefit from more detail on tobacco’s alkaloids (nicotine, nornicotine, anatabine), flavonoids and terpenes, including approximate concentrations in Nicotiana tabacum L. leaves. Response: Revised and added (Page 5, line 100-109). 6. Correct the repeated name in line 112. Response: Checked and corrected. 7. Cigarette butts contain many toxic compounds, such as arsenic, a heavy metal. Highlighting the insecticidal potential of cigarette butt-derived products could strengthen the manuscript. "What about the toxicity of the present compounds? Is it reliable and safe to use them as insecticides?". Response: We thank the reviewer for his important observation regarding the presence of toxic compounds in cigarette butt waste. Although cigarette butts contain toxic compounds such as arsenic, extracts obtained from plant waste (including tobacco waste) contain a variety of phytochemicals, including alkaloids (such as nicotine), flavonoids, terpenes, and other bioactive compounds with clear insecticidal effects. Furthermore, an assessment of the effect on non-target organisms showed that the extracts had negligible or statistically insignificant effects on most of these organisms, indicating the extracts' ability to target mosquito larvae without causing significant harm to beneficial organisms. 8. Add a sentence in the Introduction (or early Discussion) succinctly stating how cigarette butt–derived larvicides compare in cost, environmental impact, or ease of production versus conventional options. Response: Revised and added in early discussion (Page 17, line 398-401). 9. The Materials and Methods and Results sections are well-structured and clearly presented. Response: Thank you for your careful review and constructive comments. 10. Was any standard control (Pesticide) used for comparison in this study? Please clarify. Response: In our current study, unfortunately, we did not use a standard control (pesticide), because there are many biological agents that are focused on the tobacco plants and their cigarette butt waste on two vector-borne diseases: Culex pipiens and Aedes aegypti. 11. Figures 4 to 8 currently show mean values only. Add standard error or confidence interval bars to all bar charts and line plots to indicate variability. Response: Revised and added. 12. Was ethical clearance obtained for the use of hamster mouse in this study? If so, please provide the relevant details in the section 2.3.1. Response: The research title, overall objective of the study, and ethical approval for the use of hamsters in this study are presented to the Ethics Committee of the Faculty of Science, and it is approved under Code (BUFS-REC-2025-355 Ent); (Page 10, line 223-226). 13. The Discussion section is well-structured but is overly lengthy. Consider condensing or reorganizing it for better flow and add the limitation and future direction of the study. Response: Revised and Corrected. 14. The Conclusion should be rewritten. Please summarize the core findings and include future perspectives. Response: Revised and rewritten. 15. Cross check that all figures/tables are cited in order. Response: Checked and revised. 16. The entire manuscript should be thoroughly checked for grammatical, typographical, and spacing errors. Response: Checked, revised and corrected. Reviewer #3: Response: Thank you for your careful review and constructive comments. 1. Lines 26 to 33 of the abstract should be rewritten in more concise and informative way. Response: Revised and rewritten. 2. The abstract lacks the main phytochemical results. Authors should highlight the main identified phytoconstituents. Response: Checked, added. 3. In the plant collection part, kindly add the name of the botanist who authenticated the collected tobacco plant. Response: Revised and added. 4. In lines 182 and 187, why only the plant extract and one brand were selected for analysis rather than the other collected cigarette butt samples? Response: Thank you for the important note. Tobacco plant extract and one cigarette butt brand (Cleopatra) were selected for spectroscopic analysis (UV-Visible and FTIR) because these samples represent the basic chemical composition and common active compounds found in most of the cigarette butt samples, we collected. The choice was indicated based on preliminary analyses that showed a high similarity in composition between the various cigarette butt samples, which led us to believe that analysing a single sample (CBs) with the tobacco plant (N. tabacum) was sufficient to provide a clear picture of the chemical constituents of this sample class (CBs) while saving the study resources and focusing on the results of interest. Accordingly, Cleopatra was selected as a common brand and representative of the chemical composition of other cigarette butt extracts. 5. Upon collecting the cigarette butts from public areas, how could the authors differentiate the exact brand of each of them? Response: When collecting cigarette butts from public places, we relied on visual markers on filters and scrap paper to identify each cigarette pack's brand. We also relied on designs and logos printed on unfinished cigarette butts or filter pieces. Additionally, the brand identity of the collected cigarette butts was standardized and explained to individual staff working in these public places. This process helped reduce sample classification errors and ensure accurate analysis. 6. The authors are advised to check for any Grammatik or spelling mistakes throughout the manuscript. Response: Checked, corrected. 7. Line 352, the authors mistype “acetone extract” although the study is for methanol extract. correct please. Response: Revised and corrected (Methanolic), Page 16, line 378. 8. A standardization method, using HPLC for example, for the percentage of different phytochemicals present in the cigarette butt extracts is requested. Response: Thank you for the important note. We would like to clarify that in the current study, HPLC was not applied or used to analyze and standardize the phytochemical ratios in cigarette butt extracts. Instead, GC-MS, FTIR, and UV-visible spectroscopy analyses were used to identify and characterize the active chemical compounds. 9. The discussion part is too long and need to be rewritten in more informative way. Response: Checked and revised. 10. The discussion part may include the authors’ opinion about the feasibility of using cigarette butt waste as a source for bio insecticide. Response: Revised and added (page 22, line 536-550). 11. In the references part, the latin names should be italic. Response: Checked and revised. 12. In table 9, instead of bioactive compounds in the header, the authors may use chemical class or class. Response: Revised and corrected in tables 9-11. 13. In fig. 2 kindly add the scale used in capturing this picture. Response: Revised and Corrected. 14. Kindly provide an explanation for figure 10, the FTIR results, in the results section. Response: Revised and added more results about the explanation of FTIR results. Attachments Attachment Submitted filename: Response_to_reviewer_comments_Butts_Plos_R1.doc https://doi.org/10.1371/journal.pone.0333549.r002
3 Sep 2025 Decision Letter - Jorddy Neves Cruz, Editor PONE-D-25-33977R1Field and Laboratory Assessment of Larvicidal Activity of Tobacco Plants and The Cigarette Butt Waste on Culex pipiens(Linnaeus, 1758), Aedes aegypti (Linnaeus, 1762) L. and Non-Target OrganismsPLOS ONE Dear Dr. Selim, 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 Oct 18 2025 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. Please include the following items when submitting your revised manuscript: A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'. A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'. An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'. If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter. If applicable, we recommend that you deposit your laboratory protocols in protocols.io to enhance the reproducibility of your results. Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: https://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols. Additionally, PLOS ONE offers an option for publishing peer-reviewed Lab Protocol articles, which describe protocols hosted on protocols.io. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols. We look forward to receiving your revised manuscript. Kind regards, Jorddy Neves Cruz Academic Editor PLOS ONE Journal Requirements: 1.If the reviewer comments include a recommendation to cite specific previously published works, please review and evaluate these publications to determine whether they are relevant and should be cited. There is no requirement to cite these works unless the editor has indicated otherwise. Additional Editor Comments: Authors must provide a point-by-point review of all referees' queries. All modifications should be in yellow for better visualization of the manuscript changes. [Note: HTML markup is below. Please do not edit.] [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. https://doi.org/10.1371/journal.pone.0333549.r003
Revision 2
13 Sep 2025 Author Response Journal Requirements: Point#: If the reviewer comments include a recommendation to cite specific previously published works, please review and evaluate these publications to determine whether they are relevant and should be cited. There is no requirement to cite these works unless the editor has indicated otherwise. Response#; it edit Additional Editor Comments: Point#: Authors must provide a point-by-point review of all referees' queries. All modifications should be in yellow for better visualization of the manuscript changes. Response#: all comments of reviewers highlighted by yellow color Attachments Attachment Submitted filename: Response to reviewers comments.doc https://doi.org/10.1371/journal.pone.0333549.r004
16 Sep 2025 Decision Letter - Jorddy Neves Cruz, Editor Field and Laboratory Assessment of Larvicidal Activity of Tobacco Plants and The Cigarette Butt Waste on Culex pipiens(Linnaeus, 1758), Aedes aegypti (Linnaeus, 1762) L. and Non-Target Organisms PONE-D-25-33977R2 Dear Dr. Selim, 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 will be generated when your article is formally accepted. Please note, if your institution has a publishing partnership with PLOS and your article meets the relevant criteria, all or part of your publication costs will be covered. Please make sure your user information is up-to-date by logging into Editorial Manager at Editorial Manager® and clicking the ‘Update My Information' link at the top of the page. For questions related to billing, please contact billing support. 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, Jorddy Neves Cruz Academic Editor PLOS ONE Additional Editor Comments (optional): Reviewers' comments: https://doi.org/10.1371/journal.pone.0333549.r005
Formally Accepted
Acceptance Letter - Jorddy Neves Cruz, Editor PONE-D-25-33977R2 PLOS ONE Dear Dr. Selim, I'm pleased to inform you that your manuscript has been deemed suitable for publication in PLOS ONE. Congratulations! Your manuscript is now being handed over to our production team. At this stage, our production department will prepare your paper for publication. This includes ensuring the following: * All references, tables, and figures are properly cited * All relevant supporting information is included in the manuscript submission, * There are no issues that prevent the paper from being properly typeset You will receive further instructions from the production team, including instructions on how to review your proof when it is ready. Please keep in mind that we are working through a large volume of accepted articles, so please give us a few days to review your paper and let you know the next and final steps. Lastly, 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. You will receive an invoice from PLOS for your publication fee after your manuscript has reached the completed accept phase. If you receive an email requesting payment before acceptance or for any other service, this may be a phishing scheme. Learn how to identify phishing emails and protect your accounts at https://explore.plos.org/phishing. If we can help with anything else, please email us at customercare@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. Jorddy Neves Cruz Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0333549.r006

Open letter on the publication of peer review reports

PLOS recognizes the benefits of transparency in the peer review process. Therefore, we enable the publication of all of the content of peer review and author responses alongside final, published articles. Reviewers remain anonymous, unless they choose to reveal their names.

We encourage other journals to join us in this initiative. We hope that our action inspires the community, including researchers, research funders, and research institutions, to recognize the benefits of published peer review reports for all parts of the research system.

Learn more at ASAPbio .