PONE-D-24-12291Yeast encapsulation of photosensitive insecticides increases toxicity against mosquito larvae while protecting microorganismsPLOS ONE

Dear Dr. Hillyer,

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.

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ACADEMIC EDITOR: Please insert comments here and delete this placeholder text when finished. Be sure to:Reviewer 1

• In the materials and methods, the authors need to clarify few points.
  Line no. 125 How did the authors discover that pupation doesn't occur? Kindly specify. Did you do any test or Preliminary studies? is this information fr om references?
  Line no 139 How much is the light or dark time? How many lux ? Kindly add this information.
  In discussion:
  Line no 324 Pl modify this phrase. It seems incomplete.
  Line no. 350 Pl mention in materials and methods regarding microscopic observations

Reviewer 2Yeast encapsulation of photosensitive insecticides increases toxicity against mosquito larvae while protecting microorganisms

Comments

1. Line 18 - evolution and spread - evolution and subsequent spread

2. Line 23 - However, no effective delivery strategies exist – The statement is quite strong and controversial, change it into currently, there are limited effective delivery strategies...

3. The author can mention few quantitative results, in the abstract.

4. Line 57 - as seen by curcumin not being toxic against adult Danio rerio (zebrafish) – Why sudden comparison with zebrafish?

5. Justify the writing

6. Line 88 - in 4th instar – with 4th instar

7. Line 124 - because this is not a time when larvae pupate, and we discovered that pupation protects larvae from PSI toxicity – change it into a more standard form - as larvae do not pupate during this time, and we discovered that pupation protects larvae from PSI toxicity.

8. Line 100 - either 20 μM methylene blue or 100 μM curcumin were mixed in a 1000 mL Erlenmeyer flask – Why specifically was those 2 concentrations were chosen?

9. Why ethanol is used as a control? What is its significance in this work?

10. Line 200 - we first sought to determine - Therefore, we first aimed to (Keep the language in a standard research article writing tone)

11. Line 291 - does not meaningfully affect - because it does not significantly

12. Line 198 - The phototoxicity of curcumin, however, has only been investigated in Aedes aegypti larvae [7, 16, 17]. – Why only specifically to Aedes aegypti?

13. Line 269 - we suspected that larvae consume more PSIs when they are encapsulated by yeast – Assumptions are not dependable, use proper references, or any occurrence to support this statement.

14. Line 350 - It is possible that the binding of PSIs to yeast components makes them too large to diffuse across the gut epithelium – Cite this

15. Introduction lacks proper transition, from new mosquito strategy, it jumps directly to PSIs, give it a smooth flow.

16. Was there a proper control for the amount of free curcumin or methylene blue that might be released from yeast cells?

17. There’s an assumption that larvae ingest more PSI when encapsulated in yeast. Is there direct evidence like an image of gut content to support this? If yes include it.

18. Different larval stages might respond differently to PSIs. Is there any reason stage 4 were specifically selected?==============================

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Kind regards,

Muthugounder Subramanian Shivakumar, Ph.D.

Academic Editor

PLOS ONE

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Additional Editor Comments:

Dear Author

The reviewers have completed their reviews. they are suggesting a few minor corrections which needs to be carried out in the manuscript. The details of which can be found below:

Reviewer 1

In the materials and methods, the authors need to clarify few points.

Line no. 125 How did the authors discover that pupation doesn't occur? Kindly specify. Did you do any test or Preliminary studies? is this information fr om references?

Line no 139 How much is the light or dark time? How many lux ? Kindly add this information.

In discussion:

Line no 324 Pl modify this phrase. It seems incomplete.

Line no. 350 Pl mention in materials and methods regarding microscopic observations

Reviewer 2

Yeast encapsulation of photosensitive insecticides increases toxicity against mosquito larvae while protecting microorganisms

Comments

1. Line 18 - evolution and spread - evolution and subsequent spread

2. Line 23 - However, no effective delivery strategies exist – The statement is quite strong and controversial, change it into currently, there are limited effective delivery strategies...

3. The author can mention few quantitative results, in the abstract.

4. Line 57 - as seen by curcumin not being toxic against adult Danio rerio (zebrafish) – Why sudden comparison with zebrafish?

5. Justify the writing

6. Line 88 - in 4th instar – with 4th instar

7. Line 124 - because this is not a time when larvae pupate, and we discovered that pupation protects larvae from PSI toxicity – change it into a more standard form - as larvae do not pupate during this time, and we discovered that pupation protects larvae from PSI toxicity.

8. Line 100 - either 20 μM methylene blue or 100 μM curcumin were mixed in a 1000 mL Erlenmeyer flask – Why specifically was those 2 concentrations were chosen?

9. Why ethanol is used as a control? What is its significance in this work?

10. Line 200 - we first sought to determine - Therefore, we first aimed to (Keep the language in a standard research article writing tone)

11. Line 291 - does not meaningfully affect - because it does not significantly

12. Line 198 - The phototoxicity of curcumin, however, has only been investigated in Aedes aegypti larvae [7, 16, 17]. – Why only specifically to Aedes aegypti?

13. Line 269 - we suspected that larvae consume more PSIs when they are encapsulated by yeast – Assumptions are not dependable, use proper references, or any occurrence to support this statement.

14. Line 350 - It is possible that the binding of PSIs to yeast components makes them too large to diffuse across the gut epithelium – Cite this

15. Introduction lacks proper transition, from new mosquito strategy, it jumps directly to PSIs, give it a smooth flow.

16. Was there a proper control for the amount of free curcumin or methylene blue that might be released from yeast cells?

17. There’s an assumption that larvae ingest more PSI when encapsulated in yeast. Is there direct evidence like an image of gut content to support this? If yes include it.

18. Different larval stages might respond differently to PSIs. Is there any reason stage 4 were specifically selected?

[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

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

Reviewer #1: Yes

Reviewer #2: 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: No

Reviewer #2: 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: 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: In the materials and methods, the authors need to clarify few points.

Line no. 125 How did the authors discover that pupation doesn't occur? Kindly specify. Did you do any test or Preliminary studies? is this information fr om references?

Line no 139 How much is the light or dark time? How many lux ? Kindly add this information.

In discussion:

Line no 324 Pl modify this phrase. It seems incomplete.

Line no. 350 Pl mention in materials and methods regarding microscopic observations.

Reviewer #2: Yeast encapsulation of photosensitive insecticides increases toxicity against mosquito larvae while protecting microorganisms

Comments

1. Line 18 - evolution and spread - evolution and subsequent spread

2. Line 23 - However, no effective delivery strategies exist – The statement is quite strong and controversial, change it into currently, there are limited effective delivery strategies...

3. The author can mention few quantitative results, in the abstract.

4. Line 57 - as seen by curcumin not being toxic against adult Danio rerio (zebrafish) – Why sudden comparison with zebrafish?

5. Justify the writing

6. Line 88 - in 4th instar – with 4th instar

7. Line 124 - because this is not a time when larvae pupate, and we discovered that pupation protects larvae from PSI toxicity – change it into a more standard form - as larvae do not pupate during this time, and we discovered that pupation protects larvae from PSI toxicity.

8. Line 100 - either 20 μM methylene blue or 100 μM curcumin were mixed in a 1000 mL Erlenmeyer flask – Why specifically was those 2 concentrations were chosen?

9. Why ethanol is used as a control? What is its significance in this work?

10. Line 200 - we first sought to determine - Therefore, we first aimed to (Keep the language in a standard research article writing tone)

11. Line 291 - does not meaningfully affect - because it does not significantly

12. Line 198 - The phototoxicity of curcumin, however, has only been investigated in Aedes aegypti larvae [7, 16, 17]. – Why only specifically to Aedes aegypti?

13. Line 269 - we suspected that larvae consume more PSIs when they are encapsulated by yeast – Assumptions are not dependable, use proper references, or any occurrence to support this statement.

14. Line 350 - It is possible that the binding of PSIs to yeast components makes them too large to diffuse across the gut epithelium – Cite this

15. Introduction lacks proper transition, from new mosquito strategy, it jumps directly to PSIs, give it a smooth flow.

16. Was there a proper control for the amount of free curcumin or methylene blue that might be released from yeast cells?

17. There’s an assumption that larvae ingest more PSI when encapsulated in yeast. Is there direct evidence like an image of gut content to support this? If yes include it.

18. Different larval stages might respond differently to PSIs. Is there any reason stage 4 were specifically selected?

**********

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: Yashkamal K.

Reviewer #2: Yes: Chinnaperumal Kamaraj

**********

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Yeast encapsulation of photosensitive insecticides increases toxicity against mosquito larvae while protecting microorganisms

PONE-D-24-12291R1

Dear Dr. Hillyer,

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,

Muthugounder Subramanian Shivakumar, Ph.D.

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

the manuscript can be accepted

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: (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

**********

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

Reviewer #1: Yes

**********

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

**********

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

**********

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: The work is interesting and in my opinion the authors have adequately addressed the queries raised during the previous review.

**********

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: Yashkamal K

**********