PONE-D-22-26785Effects of antennal segments defects on blood-sucking behavior in Aedes albopictusPLOS ONE

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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: Yes

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

Reviewer #1: Yes

Reviewer #2: Yes

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Reviewer #1: Yes

Reviewer #2: Yes

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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: It is an interested study and provided some interested findings for further understanding the antenna with sensors /receptors function for continuing blood feeding. The results are not surprised because the antenna with sensors is for a long range purpose to find the odor and hosts. There are many studies on the host-seeking and blood feeding behaviors of Aedes albopictus in China and other Asian countries in the 1980's and 1990's. The host-seeking and blood feeding is a complicate issue, not only the receptors on the antenna, also, other parts of organs, such as papa, proborese , and tarsal, mating status, distance between hosts and mosquitoes, and other many factors influence on the host-seeking and blood- feeding behaviors. The following questions and comments may assist the authors for further revision:

1. Provide the strain of Aedes albopictus (Hunan?) and more information about the laboratory colony (how long of the colony you and they have and what kind of regular blood resource used even if this species is an opportunity feeder).

2. Have you sealed the antenna after you cut? What was the speed for orientation, probing time, and how long for the blood engorgement, compared with non-cut individual mosquitoes?

3. Provide more details about human volunteers due to the variation by individual.

4. Italic for all scientific name of the species in text and references.

5. More discussion about the receptors/sensors. There were many studies about the olfactory sensors, also, single sense cell function/role detected by EAG ..

6. There were several publications about the receptors of antenna of Aedes albopictus in China. You need to search for the Chinese literatures and understand where you stand and start. Huang, XJ, et al. 1991. Studies on the ultra-structures of antennal receptors of Aedes albopictus. Chinese Journal of Vector Biology and Control Vol. 2 (suppl.): 29-32. Xue RD. 1991. Studies on the olfactory behavior of Periplaneta americana, Musca domestica, and Aedes albopictus (NSF project summary). Chin J Vector Biol Control. Vol. 2 (suppl):5-9. Also, there is a reviewing article about this subject. Xue RD. 2016. Host-seeking and blood feeding behavior of Aedes albopictus. Technical Bulletin of the Florida Mosquito Control Association, Vol. 10:2-13. You may find this article from the ResearchGate or the Florida Mosquito Control Association website at www.yourfmca.org

Reviewer #2: Nice research, good interpretation of data. However, two points: One, this is not frontline research. I have attached a file the stand of the research of today. With the molecular tools we have today, we should use them to get a clear idea where we could interact genetically. The field is so hot these days that it is hard to keep up. However, that is the name of the game. Two, by cutting the antennae short one does cut all sensory system short. To attribute a change of behavior to olfaction may not be correct. One would have to show that mechanoreception has not been altered.

I can see that a more thorough literature review would improve the manuscript. For example, similar research has been conducted with other small animals (e.g., zooplankton, fruitflies) with similar results. This would then give a better platform for their observed data. --> major edit

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Reviewer #1: Yes: Rui-De Xue

Reviewer #2: Yes: J R Strickler

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Attachments

Attachment

Submitted filename: MOSSY-6-s13071-022-05259-x.pdf

PONE-D-22-26785R1Effects of antennal segments defects on blood-sucking behavior in Aedes albopictusPLOS ONE

Dear Dr. Chen,

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 Apr 15 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.

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'.
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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,

Jiang-Shiou Hwang, Ph.D.

Academic Editor

PLOS ONE

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Comments to the Author

1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation.

Reviewer #2: (No Response)

Reviewer #3: (No Response)

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2. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #2: Yes

Reviewer #3: Partly

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

Reviewer #2: N/A

Reviewer #3: I Don't Know

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4. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #2: Yes

Reviewer #3: Yes

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5. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #2: Yes

Reviewer #3: No

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6. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #2: So, here my evaluation of the revised manuscript.

All little points are fine, the English is somewhat charming but understandable.

The value of this manuscript is so/so. Very old-fashioned research when compared with the research done on Drosophila, for example.

Here an abstract of similar research:

Functional Interaction Between Drosophila Olfactory Sensory Neurons and Their Support Cells

by Sinisa Prelic et al --- https://doi.org/10.3389/fncel.2021.789086

Insects detect volatile chemicals using antennae, which house a vast variety of olfactory sensory neurons (OSNs) that innervate hair-like structures called sensilla where odor detection takes place. In addition to OSNs, the antenna also hosts various support cell types. These include the triad of trichogen, tormogen, and thecogen support cells that lie adjacent to their respective OSNs. The arrangement of OSN supporting cells occurs stereotypically for all sensilla and is widely conserved in evolution. While insect chemosensory neurons have received considerable attention, little is known about the functional significance of the cells that support them. For instance, it remains unknown whether support cells play an active role in odor detection, or only passively contribute to homeostasis, e.g., by maintaining sensillum lymph composition. To investigate the functional interaction between OSNs and support cells, we used optical and electrophysiological approaches in Drosophila. First, we characterized the distribution of various supporting cells using genetic markers. By means of an ex vivo antennal preparation and genetically-encoded Ca2+ and K+ indicators, we then studied the activation of these auxiliary cells during odor presentation in adult flies. We observed acute responses and distinct differences in Ca2+ and K+ fluxes between support cell types. Finally, we observed alterations in OSN responses upon thecogen cell ablation in mature adults. Upon inducible ablation of thecogen cells, we notice a gain in mechanical responsiveness to mechanical stimulations during single-sensillum recording, but a lack of change to the neuronal resting activity. Taken together, these results demonstrate that support cells play a more active and responsive role during odor processing than previously thought. Our observations thus reveal that support cells functionally interact with OSNs and may be important for the extraordinary ability of insect olfactory systems to dynamically and sensitively discriminate between odors in the turbulent sensory landscape of insect flight.

Reading this abstract we can see that the discussion about "wound healing" is way out considering what the researchers above published.

In this context, the results of the authors are fine as results. One has to start somewhere. However, they have to connect with literature like above to make it clear, that their research is the first step to more targeted research where one would use their step as step one and continue with research as above as the guiding research for further meaningful research.

I realize that this field is very competitive and to get a clear step forward is hard to achieve.

So, based on this, it is your decision.

One, the research is not up to the scientific level of your journal;

or

Two, for a review one would need another round of editing with a clear appreciation of the research done on other species (see literature cited in above).

I would prefer solution TWO. These researchers would gain a lot of new knowledge and would move in the direction of modern olfactory research. However, you may not have the pages for it (--> send it to an appropriate journal).

Reviewer #3: The investigation has been done on very obvious and general objective: whoever study basic moth part and feeing mechanisms can expect antenatal segment efects will affect blood sucking behaviour. However, the specific study on mosquito antennal flagellomeres and particularly is an enhancement of nowledge. Which flagellomere may affect their blood-feeding behavior, and through rationally designed behavioral experiments, we found that the 6th and 7th flagellomeres on Aedes albopictus antenna are important in the olfactory detection of host searching. Meanwhile, the morphology and distribution of sensilla on each antenna flagellomere were also analysed and discussed in this paper. Over all It is an interesting study and provided some new findings for further understanding the antenna with sensors /receptors function. However, the results are nothing rather, along the expected line.

My specific comments are as follow:

Abstract: Too general statement has been provided in the introductory sentences in abstract The blood-feeding transmits extremely harmful infections including malaria, yellow fever, dengue fever, and other arboviruses, making mosquitoes one of the most harmful creatures to human health.

Methods:

Authors have neither performed proper sealing of the antenna, nor studied orientation activity of the antenna, this implication some line of future studies should be explained.

Repetition of the scientific name Aedes albopictus

Write ones’ full name then abbreviate form as Ae albopictus

Discussion: There should be some lines of discussion on the host-seeking and blood feeding behaviour as this is a complicated issue, not only the receptors on the antenna, also, other parts of organs, such as proborese , and tarsal, mating status, distance between hosts and mosquitoes, etc. are also important determinant of the behaviour so proper discussion explaining implication of these results on vector transmission, blood sucking an knowledge enhancement behavioural attributes of host seeking blood feeing should be explained.

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Reviewer #2: No

Reviewer #3: No

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PONE-D-22-26785R2Effects of antennal segments defects on blood-sucking behavior in Aedes albopictusPLOS ONE

Dear Dr. Chen,

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 27 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.

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'.
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• 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.

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We look forward to receiving your revised manuscript.

Kind regards,

Jiang-Shiou Hwang, Ph.D.

Academic Editor

PLOS ONE

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Please review your reference list to ensure that it is complete and correct. If you have cited papers that have been retracted, please include the rationale for doing so in the manuscript text, or remove these references and replace them with relevant current references. Any changes to the reference list should be mentioned in the rebuttal letter that accompanies your revised manuscript. If you need to cite a retracted article, indicate the article’s retracted status in the References list and also include a citation and full reference for the retraction notice.

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Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation.

Reviewer #2: All comments have been addressed

Reviewer #3: All comments have been addressed

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2. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #2: Yes

Reviewer #3: Yes

**********

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

Reviewer #2: Yes

Reviewer #3: 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 #2: Yes

Reviewer #3: 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 #2: Yes

Reviewer #3: 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 #2: Yes, do catch up with today's olfaction research. Your starting point is fine. However, for progress we need more detailed information that we can compare with results from other insects.

Reviewer #3: Revision has been done . quality of the Ms is better and understandable now. However the manuscript need minor revision before acceptance . Following clarification amendment are required

1. The revised version has an additional author hope the procedure of authorship modification has been followed.

2. The introductory sentences in abstract The blood-feeding transmits extremely harmful infections including malaria, yellow fever, dengue fever, and other arboviruses, making mosquitoes one of the most harmful creatures to human health. Is too general statement and may be deleted from abstract?

3. Introduction: line 21-24: After mating, female mosquitoes.…. blood feeding. Shorten the sentence.

4. line 41: This paper attempts…molecular level; Could be rewritten as This paper combines morphological and behavioural studies to determine the functional regions of olfactory organ of mosquitoes to narrow down the range of research targets at cellular and molecular levels.

5. Material and methods: Line 136-140; The physical characteristics of volunteer could be depicted as pointers like; Age: 33, Sex: female, Height: 165cm.. and so on.

Was there only one participant out of the 23 sign ups?

6. Line 158-159: Grammatical error; Participants were aware of the content and purpose of the study but were unaware of the test order.

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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 #2: No

Reviewer #3: No

**********

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Effects of antennal segments defects on blood-sucking behavior in Aedes albopictus

PONE-D-22-26785R3

Dear Dr. Chen,

We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements.

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