PONE-D-22-31911Optimization of rice spacing density and fish species to improve production of both rice and fish through co-culture systemPLOS ONE

Dear Dr. Inayat,

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: I have critically gone through the manuscript  and the Ms reviewed by two very efficient reviewers working in the field of capture and culture fishery. Please go through those comments and reconsider to make title more appealing and directly addressing objectives.   Both the reviewers have similar views and I also consider required revision required along the same line. The manuscript has neither page number nor line number, so I find difficulty in commenting objectively.

The Ms. has eight tables and three figures,

As zooplankton and phytoplankton community are not main goal of the study I would suggest to shift these tables to supplementary tables (Figure 4 and 5). However zooplankton like cyclopoid copepods are abundantly found in race paddy field, many species are predators of fish larvae. Do they affect fish production in rice paddy field. This issue may be brought in discussion with references and author's zooplankton data.

For Lab, Study and Registered Report Protocols: These article types are not expected to include results but may include pilot data. 

==============================

Please submit your revised manuscript by Feb 04 2023 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file.

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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,

Ram Kumar, Ph.D.

Academic Editor

PLOS ONE

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2. Thank you for stating the following financial disclosure:

“Yes.

This research work was funded by Punjab Agriculture Research Board (PARB), Punjab, Pakistan under project no. 674.”

Please state what role the funders took in the study.  If the funders had no role, please state: "The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."

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Please include this amended Role of Funder statement in your cover letter; we will change the online submission form on your behalf.

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We note that you have provided funding information that is currently declared in your Funding Statement. However, funding information should not appear in the Acknowledgments section or other areas of your manuscript. We will only publish funding information present in the Funding Statement section of the online submission form.

Please remove any funding-related text from the manuscript and let us know how you would like to update your Funding Statement. Currently, your Funding Statement reads as follows:

“Yes. 

This research work was funded by Punjab Agriculture Research Board (PARB), Punjab, Pakistan under project no. 674.”

Please include your amended statements within your cover letter; we will change the online submission form on your behalf.

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Additional Editor Comments:

The manuscript has neither page number nor line number, so I find difficulty in commenting objectively.

The Ms. has eight tables and three figures,

As zooplankton and phytoplankton community are not main goal of the study I would suggest to shift these tables to supplementary tables (Figure 4 and 5). However zooplankton like cyclopoid copepods are abundantly found in race paddy field, many species are predators of fish larvae. Do they affect fish production in rice paddy field. This issue may be brought in discussion with references and author's zooplankton data.

Tables 7 and 8 can be nicely presented as graph as simple vertical bars, X axis: parameters and Y axis: Values (pH values may be given on secondary Y axis or omitted

Proper proof reading has not been done carefully, e.g. Legend of table 1 is given below that table where as legend of table 2, 3 are given above tables. Similarly Figure 2 is inserted within the text but other Figures are given at the end of text, after reference section

The statement appendix of tables as "Superscripts on different means within row differ significantly at P ≤ 0.05; RHD= Rice high 8 density (RHD), RMD= Rice medium density (RMD), RLD= Rice low density (RLD), is not clearly communicate the message what (i) if two means are superscripted by same alphabets and (ii) if two means are superscripted by different alphabets ??

Figure 3: What is purpose of line drawing of values, which are not continuous and different points represent different parameters .

All sections of the manuscript need restructuring and rewriting. Language is verbose.

A completely rewritten manuscript focusing on objectives of the study may be considered for another round of assessment.

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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

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

Reviewer #1: Yes

Reviewer #2: I Don't Know

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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: No

Reviewer #2: 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: No

Reviewer #2: 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: Journal; PLOS ONE

Ref No.: PONE-D-22-31911

General comments:

The present research work has no such significant importance and advancement in this field. There is no such novelty in the present study, since this types of study has already been performed.

Specific comments:

Title: Title should be more clear. Please rrewrite title

Abstract: Rewrite it clarifying the following comments

1. The abstract is not self-explanatory.

2. It should be more specific and cclear

3. Where is the results of rice?

4. Novelty statement?

Introduction:

1. It is very good and informative.

2. Please insert some relevant current references from other authors if available.

3. “one word, from the perspective of fish and rice, the optimum rice spacing density and optimum fish species for the culture of rice and fish have remained silent.” Is it right ? There are several studies.

4. Need more current references.

5. Rice spacing, density .. is not proper

Materials and methods

It is not well structured. Need to separate the rice, fish parameters, water soil parameters

Experimental design should contain fish stocking

It is ok.

Results

It is not well structured. Presents the results on the basis of important results. Rice and fish production parameters should come first.

Need to rewrite it properly.

Only table, need some graphical presentation.

Discussion

I think author should deal with the result-wise (parameter-wise) outcome of the present study supporting previous and current referential study. Need more specific discussion without general statements as mentioned in the starting of the discussion. Need more analytical discussion on the basis of results obtained from study

Insert a clear concluding sentence with your significant supporting evidence obtained.

Table: Ok. Check caption properly.

References: Not checked. Please check the references.

Reviewer #2: Comments- The manuscript requires rigorous re-editing and proof reading, to ensure clarity. The research work is very common, and using advanced tools to support the data obtained would greatly benefit the research area, and add novelty.

The title should include the key species used for the experiment.

The abstract should provide proper insight about the research, with more clarity.

GMT, RMD, RLD, RHD, are mentioned several times, before the first mention of their unabbreviated form.

Maintain uniformity in the use scientific name, or common name.

The introduction is informative, and could be re-written to make it better.

Add more recent references for the general statements. There are very few recent references used in the manuscript.

Materials and methods

Is "description of the field" appropriate enough for the specifications of the study area?

How is the climate, elevation, and temperature relevant to the research conducted?

Numerous grammatical errors are present, and the section could be restructured.

Fig 2, the symbols used after A,B, and C, do not match the ones in the figure.

Where were the fishes procured from? Were they acclimatized prior to use in the experiment? If yes, then in what conditions. If no, why not? What was the age of the fishes used in the experiment.

How were the feeding and growth conditions accessed, to increase the quantity of the feed?

Is "evaluation of meat analysis" the best fit ?

How were the zooplankton identified?

The materials and methods should be described clearly enough for the work to be reproducible.

Results

Re-write it more clearly.

Some of the scientific names mentioned are italicized, without capitalizing the first word. Some are written with no space between the generic name and specific name.

The most important results of the study are related to the rice and fish, that should be mentioned first.

Discussion

The discussion has too many general statements, better suited for introduction. Add more scientific discussion and interpretation, relevant to the results obtained.

Alanine, Glycine and such amino acids can be mentioned in their full name, before using Ala, Gly etc.

Re-write, with rigorous proof -reading, to remove elementary level grammatical errors.

Check PLOS one guidelines are formatting for references and change accordingly.

Table

Use "Species" instead of "Specie"

Italicize the scientific names.

What are "Ostracodans"?

"Protozoan", "Cladocera" , "Copepods" , "Rotifers" maintain uniformity.

Is "Bosmiina" the correct word?

Why are there line numbers from table 4?

What is the "f" in Line 60

Add line numbers to the entire manuscript, instead of the Table.

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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

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Attachments

Attachment

Submitted filename: Paper review PLOS.doc

PONE-D-22-31911R1

Optimizing Rice-Fish Co-Culture Systems: Investigating the Impact of Spacing Density and Fish Species Selection on Bio-Chemical Profile and Production

PLOS ONE

Dear Dr. Inayat, 

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we have decided that your manuscript does not meet our criteria for publication and must therefore be rejected.

Specifically: The reviewers have serious objection on methods used and data collected. The study is not novel as large number of similar studies on same objectives and better experimental design has been published. 

I am sorry that we cannot be more positive on this occasion, but hope that you appreciate the reasons for this decision.

Kind regards,

Ram Kumar, Ph.D. D. Sc.(H/C)

Academic Editor

PLOS ONE

Additional Editor Comments (if provided):

We are sorry, for this decision but comments and suggestions made by reviewers will be useful for future work.

[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: (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 #2: Partly

**********

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

Reviewer #2: 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

**********

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: 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: 1. The authors have stated in their response that the research work is the first study of its kind, worldwide, while there are several studies accessing the impact rice spacing densities and fish species in such systems.

2. The abstract omits the bio-chemical aspect of this research work, and is not self-explanatory.

3. L99-L102. How is the climate, elevation, and temperature relevant to the research conducted? An explanation is essential why the above factors may influence the research results.

4. L163, The protocol for evaluation of amino acid profile should be mentioned in detail, with the mention of any specific standardisation carried out, if any.

5. How were the zooplankton identified, after detecting them under the microscope?

6. The manuscript would benefit from re-structuring the statements, and rigorous proof reading.

**********

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: No

**********

[NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.]

- - - - -

For journal use only: PONEDEC3

PONE-D-22-31911R2Optimizing Rice-Fish Co-Culture Systems: Investigating the Impact of Spacing Density and Fish Species Selection on Bio-Chemical Profile and ProductionPLOS ONE

Dear Dr. Inayat,

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

• Indicate which changes you require for acceptance versus which changes you recommend
• Address any conflicts between the reviews so that it's clear which advice the authors should follow
• Provide specific feedback from your evaluation of the manuscript

Please ensure that your decision is justified on PLOS ONE’s publication criteria and not, for example, on novelty or perceived impact.

For Lab, Study and Registered Report Protocols: These article types are not expected to include results but may include pilot data. 

==============================

Please submit your revised manuscript by Sep 21 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'.
• 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,

SSS Sarma

Academic Editor

PLOS ONE

Journal Requirements:

1. Thank you for stating the following financial disclosure:

"Yes. This research work was funded by Punjab Agriculture Research Board (PARB), Punjab, Pakistan under project no. 674. "

Please state what role the funders took in the study. If the funders had no role, please state: "The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."

If this statement is not correct you must amend it as needed.

Please include this amended Role of Funder statement in your cover letter; we will change the online submission form on your behalf.

Additional Editor Comments (if provided):

Dear Authors

Please revise your ms based on fresh comments made by the reviewers. Plus you will also find my observations for improving your presentation.

I have checked the manuscript too. There is some justification that manuscript is not yet ready as pointed out by the reviewers. I would like to give one more opportunity to the authors to revise their contribution and submit it to journal for possible consideration.

The following additional aspects need to be taken while re-submitting a revised version:

1. Delete all p values from the Abstract. It is sufficient to say that significant differences exist among treatments.

2. In the title please provide the taxonomical names fish species and the authority of the taxon. This ensures proper identification of the taxa

3. Last sentence of the abstract change as “The study provides data to understand…

4. Lines 55-56: Many parasites from the field reach the ducks, fish etc. Mention necessary limitations in the conclusions about this

5. The pond length for cultivation of rice is shallow but for culturing fish species, higher depth is needed. Make necessary justification

6. It is necessary to use post hoc tests following anova. In statistical analysis some anomalous trends have been depicted. For example, Table 1 GMT weight gain in Tilapia and carp the differences shown as significant due to different letters of superscript. This requires confirmation because standard deviation is very large. In the same table, data were presented in real units and in percentage or per day. This is considerable duplication of information. First column is sufficient.

7. This applies to other tables too.

8. I have to fully checked all the tables and figures. But it is important to avoid presenting the same data in figures and tables.

9. Provide results of multiple comparisons for the bars of figs. 3-5 (with alphabets).

10. I failed to understand why the discrete bars are connected by a dotted line. This gives the impression that continuous data were collected. This is used for variables like temperature.

11. Do not start a sentence with a number (line 351).

12. The ms has 71 references. Weed out excessive refs and restrict to 50 citations. Fig. 1 showing the world map for Pakistan is too basic. Remove this blue square fig.

13. A revised version may or may not be sent out for additional reviewing.

Handling Editor

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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 #1: (No Response)

Reviewer #2: (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 #1: Partly

Reviewer #2: No

**********

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

Reviewer #1: Yes

Reviewer #2: 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 #1: Yes

Reviewer #2: 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: No

Reviewer #2: 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 #1: Comments:

The title of MS is still not clear.

The write up of MS is still not standard.

For example:

Fat % = Fat weight (g)/weight if sample ×100

Before the harvesting of rice, rice stem characteristics were measured (Fig 3). This sentence can be of M & M part.

Crude protein % = Volume ×0.875/sample weight

Need references for the methods used in study for Yield parameter of rice

Under different rice spacing density, least square mean ± standard errors of crude protein

and fat of the muscles of the fish are presented in

Figures axis is invisible.

Reviewer #2: The previous comments to the manuscript should have been incorporated in the resubmitted version.

It was strongly suggested that the proof reading be done rigorously, but the current manuscript has many errors which could have been solved with proper proof reading. A few examples are as follows-

L20- “Genetically Male Tilapia” has already been mentioned to be referred with “GMT”, yet L25, L27 mention “GMT(Tilapia)”

L26- “Delicious” is not the proper adjective to use for amino acids, in a scientific research article .

L28- Full stop/Period between “acid categories” and “with RMD”

L45, L46- Use of “and” twice in the sentence.

L58- Use of “IAAF” and then “IAAFS” for Integrated Agri-Aquaculture Farming System

L58- “IAAFS” is not required to be in bracket.

L60- Grammatical error- “improves” instead of “improve”

L66-L68- The whole sentence needs restructuring to remove grammatical errors

L168- Both curly bracket and square bracket are used at once.

L152- “Meat analyses” is very unclear, the specific tissues to be analyzed are to be mentioned.

The errors of the above mentioned kind, exist too frequently in the manuscript.

2. The authors have stated that information on climate, elevation and temperature of the study area is essential to interpret the results of the study, yet there is no interpretation on that aspect in the discussion. There should also be a proper introduction for how these factors can be helpful in interpreting the results.

3. The experimental design states that, the study is conducted on 12 plots of 6500sq.ft each, fishes were stocked at a rate of 9000/ha, meaning approximately total 6500 fishes were stocked , of which only 3 fishes per plot were sampled for proximate and amino acid analysis? Were the fishes used in proximate analysis and Amino acid analysis same? A sample size of three fishes per plot is too small, the number of replicates should be more.

4. The experimental design is not clear.

5. The authors have removed all information about zooplankton identification, but have responded to my previous query, hence I would like to mention that it is important to cite the taxonomic key and reference material used for zooplankton identification, as it not only helps the author to validate their work during the peer- review process, but also helps the reader to conduct similar study when the manuscript is published.

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Reviewer #1: No

Reviewer #2: No

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PONE-D-22-31911R3Optimizing Rice-Fish Co-Culture: Investigating the Impact of Rice Spacing density on Biochemical Profiles and Production of Genetically Modified Tilapia (Oreochromis spp.) and Cyprinus carpioPLOS ONE

Dear Dr. Inayat,

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 Dec 29 2023 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file.

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We look forward to receiving your revised manuscript.

Kind regards,

SSS Sarma

Academic Editor

PLOS ONE

Journal Requirements:

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.

Additional Editor Comments:

Dear authors

Your re-reviewed ms has come back with addition corrections. This is due to the fact that the previously two reviewers declined to take up your ms for further reviewing. As I see some potential in your ms, I have decided to give you another fair chance revise it further. Please note that your next revised ms (R4) will not be reivewed again. I will check it and send my observations to the Editor in Chief for his/her final decision. To increase the chances of positive consideration of your ms, please carefully revise your ms based on the observations of the two new reviewers. Plus please check the ms for grammatical, structural and/or typos.

Sincerely

Handling Editor

[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 #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 #3: Partly

Reviewer #4: Partly

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

Reviewer #3: N/A

Reviewer #4: Yes

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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 #3: Yes

Reviewer #4: 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 #3: Yes

Reviewer #4: 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 #3: 1. The manuscript has some new ideas and/or data.

2. The authors are interested in rice production together with fish farming

3. The authors have considered the required variables of both these groups

4. Proximate composition of different elements has been considered.

The manuscript has some weak aspects which may be addressed:

1. Discuss the role of temperature on the differential fatty acid composition.

2. The authors showed higher crude protein and fat content in genetically male tilapia tan Cyprinus carpio. Since fat composition is influenced by temperature, authors are advised to discuss this aspect

3. Fish growth depends on plankton quality and quantity. Did the authors have any idea of these aspects in different treatments?

4. The fish species excrete considerable quantities of phosphorus and nitrogen compounds which influence the plankton dynamics.

5. Did you measure the N:P ratios during the study period.

Reviewer #4: Optimizing Rice-Fish Co-Culture: Investigating the Impact of Rice Spacing density onBioch emical Profiles and Production of Genetically Modified Tilapia (Oreochromis spp.) and Cyprinus carpio

In general, the idea of the manuscript is of relevance due sustainability purposes. The evaluation of the impact of rice spacing density on the production of GMT and Cyprinus carpio is understood, however, the ideas in some sections are hard to understand. Authors may consider rewriting and revising again the editing and translation of certain concepts. Also, there are a lot of errors on spaces and end points. General trends in results are lacking. The discussion needs to be the discussion must be strengthened and conclusive, the importance of the general aim must be highlighted. Some of the references used in introduction and discussion must be updated.

Abstract.

GMT- specify

Line 73. Aquaculture

Line 80. Herbicides. Several

Line 83. friendly. In

Line 89-90. Please revise the writhing

Line 93-95. The idea is not completely comprehensible

Line 96-98 unite ideas

Line 108. There are two points

Line 111-112 authors may consider eliminating the idea, as the aim is established further on the document. Perhaps authors can consider adding the hypothesis of the study.

Line 112 The study aims to evaluate the integration…

Line 130. Eliminate propose. For the present assessment…

Line 131-132 The idea is poorly understood

Is there any reference used for the experimental design?

Line 146 study. The acclimatization…

Line 148, 151, 157 double space

Line 167 the idea is vague, specifically the ending part of the idea.

References of the keys and manuals must be added in sample collection

Line 180 fish

Line 180-Previous to the experiment

Rewrite the idea

Is there any reference of the formula? Previous study were it has been used before?

Line 188 eliminate the

The monitoring of water quality is vague. Which parameters?

Line 232. One way anova?

Line 233-235 rewrite the idea

Line 278 are presented

Line 290 were presented?

In the section of results, the general idea must be established and described. Line 298-299 is out of line.

Describe Plankton community trends

Line 311- according to what?

Line 326- according to what authors or compared to previous studies?

No trends of the results are considered in the discussion as well as the use of tables or figures.

Line 341-342 the authors may consider mentioning why…

Line 359 elaborate idea

Fig 1 modify the map due to its poor quality

Fig 2 Experimental design considering 12 plots ()

Fig 3. Standard deviation of how many replicas?

Table 1 and 2 describe the treatments groups in ()

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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 #3: No

Reviewer #4: Yes: Brenda Karen González Pérez

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[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.]

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Optimizing Rice-Fish Co-Culture: Investigating the Impact of Rice Spacing density on Biochemical Profiles and Production of Genetically Modified Tilapia (Oreochromis spp.) and Cyprinus carpio

PONE-D-22-31911R4

Dear Dr. Inayat,

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,

SSS Sarma

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

The final decision will be communicated to you by the Editorial Office.

Reviewers' comments: