PONE-D-20-21866

Use of a filamentous green alga (Chaetomorpha sp.) and microsnail (Stenothyra sp.) as feed at an early stage of intensive aquaculture promotes growth performance, artificial feed efficiency, and profitability of giant tiger prawn (Penaeus monodon)

PLOS ONE

Dear Dr. Tsutsui,

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 Nov 12 2020 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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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,

Mahmoud A.O. Dawood, PhD

Academic Editor

PLOS ONE

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 [The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.].

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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: Partly

Reviewer #2: Partly

Reviewer #3: Yes

Reviewer #4: Yes

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

Reviewer #1: Yes

Reviewer #2: No

Reviewer #3: No

Reviewer #4: Yes

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

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

Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: Yes

Reviewer #4: Yes

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

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

Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: Yes

Reviewer #4: No

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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: Author used Chaetomorpha sp. and Stenothyra sp. as feed at an early stage of intensive aquaculture of giant tiger prawn. However, there are still some problems should be improved, especially the experimental method and calculation. The following questions should be answered before further submission.

Abstract: It is hard to get to know the main content of this manuscript form the abstract. Author should provide more effective information in concise sentences.

Line 27-31: Confused. Please rewrite it.

Line 32-37: Rewrite. More results should be exhibited in abstract.

Introduction:

Line 69: References are needed here.

Line 75-82: Redundant content. Why you choose Chaetomorpha sp. and Stenothyra sp.?

Line 84-85: Why culture both Chaetomorpha sp. and Stenothyra sp. as supplementary live feeds? Why use them as feed at an early stage of intensive aquaculture?

Materials and methods:

Line 120: Why choose these amounts?

Line 121-123: How to remove other benthic organisms?

Line 123-124: How about the daily management?

Line 134: Only 20 shrimps? Compared with the initial number of shrimps, the measured quantity is quite small, whether it leads to a big error?

Discussion：

Line 280-283: Confused. Please explain these sentences.

Table 1: How do you calculate the apparent benthos consumption?

Table 3: There are no Chaetomorpha sp. Stenothyra sp. provided after 8 weeks. Why the higher feed consumption was found in experimental treatment?

Table 4: 0 may not possible. Please explain.

Table 5: What is the amount of benthos? What is the amount of benthos? Line 120: 6.81 kg and 1.96 kg. Survival rate>100%???

Reviewer #2: Apparently the experimental diet stimulated growth; the performance of P. monodon appears to have improved in the 3 groups given supplemental feeds as PLs as compared with controls. This comparison is limited statistically by the small number of groups (N = 3) and apparent variation in food consumption and survival. For these reasons I suggest that this be correctly described as as a pilot or preliminary observation and not presented as a definitive analysis. When an experimental treatment appears to hold the promise of increasing commercial production and improved profitability for a valuable crop, is is reasonable to test it on a scale that is adequate for the use of rigorous statistical analysis.

Was any cost or value ascribed to the green algae and snails that were used as supplemental feeds in this study? It is not clear to me if these organisms would be available to scale up for culture on a commercial scale. The experimental group ate more of the prepared commercial feed, so at least one mechanism of action was an increased appetite in response to the early supplemental feeds.

The discussion of IMTA on page 18 is superfluous; as the authors state, this study is not an example of integrated aquaculture, it is the addition of live feeds.

The abstract refers to the culture of “an early stage” of P. monodon; it would be preferable to identify them specifically as P. monodon postlarvae.

Line 217 Penaeus monodon should be italicized for correct scientific name format.

The different survival rates with such a low number of samples (3 control and 3 experimental ponds) is troubling – as the authors mentioned more than once, this experimental design does not allow for strong and definitive statistics. With such a small number of samples, random variation or tank effects can not be ruled out.

Reviewer #3: Table 1. check your value.

If 16.0 ± 6.1 is correct, it might be strongly possible that the statistics also are subject to change. 6.1 is high so I didn't see a statistical difference between 16.0 and 18.2.

Figures.

The figures are low resolution, which makes their observation difficult. I suggest increasing the resolution.

Reviewer #4: The authors are encourage to address the following issues:

1- The article language needs edition.

2- Abstract needs restructuring. It contains much background data and less methodological data and results. Revise it.

3- It is not clear if the added live food reproduced in the culture tank during 15 weeks of experiment. Clarify it.

4- In table 1, "total shrimp production": Unit is confusing. kg shrimp per tank? per squared meter?... Moreover, add feed intake as percentage of body weight.

5-

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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.

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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 #1: No

Reviewer #2: No

Reviewer #3: Yes: Sevdan YILMAZ

Reviewer #4: 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.

PONE-D-20-21866R1

Use of a filamentous green alga (Chaetomorpha sp.) and microsnail (Stenothyra sp.) as feed at an early stage of intensive aquaculture promotes growth performance, artificial feed efficiency, and profitability of giant tiger prawn (Penaeus monodon)

PLOS ONE

Dear Dr. Tsutsui,

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 Jan 07 2021 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: http://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols

We look forward to receiving your revised manuscript.

Kind regards,

Mahmoud A.O. Dawood, PhD

Academic Editor

PLOS ONE

Additional Editor Comments (if provided):

Authors are recommended to address the comments of reviewer 4 that has been raised in the first round of review as following:

1- The article language needs edition.

2- Abstract needs restructuring. It contains much background data and less methodological data and results. Revise it.

3- It is not clear if the added live food reproduced in the culture tank during 15 weeks of experiment. Clarify it.

4- In table 1, "total shrimp production": Unit is confusing. kg shrimp per tank? per squared meter?... Moreover, add feed intake as percentage of body weight.

Also, do you have response to the concern raised by reviewer 2?

[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 #2: All comments have been addressed

Reviewer #3: All comments have been addressed

Reviewer #4: (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: Yes

Reviewer #4: Partly

**********

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

Reviewer #2: Yes

Reviewer #3: Yes

Reviewer #4: I Don't Know

**********

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

Reviewer #4: 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

Reviewer #4: No

**********

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: The authors have argued that this study with N=3 samples is statistically valid because of the mangnitude of differences related to treatment vs control. They have made a good case statistically, and the contribution is in an acceptable form, and other comments in my initial review have been addressed satisfactorily. I am inherently uncomfortable with such small sample sizes and have done farm-scale culture studies with an N of nine, on nine commercial farms, which in my view is preferable and more convincing than making a case with a study on such a small scale.

Reviewer #3: The manuscript entitled " Use of a filamentous green alga (Chaetomorpha sp.) and microsnail (Stenothyra sp.) as feed at an early stage of intensive aquaculture promotes growth performance, artificial feed efficiency, and profitability of giant tiger prawn (Penaeus monodon) " is a well written work; the topic is very interesting for journal reader. I have reviewed revised manuscript. Many parts of revision seemed to be adequate.

Therefore, I judged this manuscript would be acceptable.

Reviewer #4: (No Response)

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7. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files.

If you choose “no”, your identity will remain anonymous but your review may still be made public.

Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy.

Reviewer #2: Yes: Christopher L. Brown

Reviewer #3: No

Reviewer #4: 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.

Use of a filamentous green alga (Chaetomorpha sp.) and microsnail (Stenothyra sp.) as feed at an early stage of intensive aquaculture promotes growth performance, artificial feed efficiency, and profitability of giant tiger prawn (Penaeus monodon)

PONE-D-20-21866R2

Dear Dr. Tsutsui,

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,

Mahmoud A.O. Dawood, PhD

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

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 #4: (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 #4: Yes

**********

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

Reviewer #2: Yes

Reviewer #4: 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 #4: 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 #4: 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: This is my third review of this manuscript, and I was already satisfied with the revisions included in the second one. Again, the number of replicate samples is very small but the statististical significance appears to be satisfactory.

Reviewer #4: The authors revised the manuscript and it is acceptable. I recommend it for pulication///////////////

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7. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files.

If you choose “no”, your identity will remain anonymous but your review may still be made public.

Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy.

Reviewer #2: Yes: Chrisotpher L. Brown

Reviewer #4: No