PONE-D-21-01652

Enhancing the quality control for mass rearing of Trichogramma evanescens Westwood reared on the Indian meal moth Plodia interpunctella (Hübner) and the wax mothGalleria mellonella L.

PLOS ONE

Dear Dr. Hasan,

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 by the two reviewers.

Please submit your revised manuscript by Apr 24 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.

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We look forward to receiving your revised manuscript.

Kind regards,

Nicolas Desneux

Academic Editor

PLOS 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: Partly

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

Reviewer #1: No

Reviewer #2: No

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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

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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

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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: The egg parasitoid Trichogramma evanescens is widely distributed throughout the world, and considered as an efficient biological control agent against many lepidopteran pests. In this study, the authors investigated effects of host species, egg age, cold storage and temperature on the quality control in mass rearing of the parasitoid. The results provide some valuable information on instructing mass production of T. evanescens with commonly intermediate hosts, the Indian meal moth, Plodia interpunctella and the wax moth, Galleria mellonella. Through checking this manuscript, the experiment design and writing is ok, the results also can support the conclusion. However, there are serious defects in data analyses. In addition, too many old references were cited in this paper. Some specific comments see the followings.

Lines 57-59: For the references of 1-7, some are too old, the authors may replace them with some new publications. Some references are recommended as followed.

Wang Y, et al., 2020. Manually-extracted unfertilized eggs of Chinese oak silkworm, Antheraea pernyi, enhance mass production of Trichogramma parasitoids. Entomologia Generalis, 40:397-406.

Zhang JJ, et al., 2018. Advantages of diapause in Trichogramma dendrolimi mass production via eggs of the Chinese silkworm, Antheraea pernyi. Pest Management Science, 74: 959-965.

Lines 60-62: The same comments to lines 57-59. Some references are recommended as followed.

Hou YY, et al., 2018. Effect of oriental armyworm Mythimna separata egg age on the parasitism and host suitability for five Trichogramma species. Journal of Pest Science, 91: 1181–1189.

Zhang JJ, et al., 2014. Effects of host-egg ages on host selection and suitability of four Chinese Trichogramma species, egg parasitoids of the rice striped stem borer, Chilo suppressalis. BioControl. 59: 159-166.

Lines 120-122: Change T.evanescens and P.interpunctella to T. evanescens and P. interpunctella.

Lines 130- 135: How long the host eggs were expose to parasitoids?

Lines 178-179: these descriptions are not matched with the experimental contents.

Lines 181-187: The data analyses are described too simple. In this paper, the authors conducted three experiments, in which different factors have been considered. The authors should describe ANOVA with multi-factors in detail.

Lines 191-192: I’m confused on df=5, 30, why the treatment freedom is 5?

Lines 194: why the treatment freedom is 3?

Lines 195-196: Total treatments are 12, why the treatment freedom is 13? All the analysis data given in the text should be checked, particular for the freedom data!

Line 207: change dold to d old.

Line 212: change 4 d to 14 d.

Lines 225-228: I can’t find the parasitism data in Table 2.

Lines 277-278: only two species, why the treatment freedom is 7? Carefully check all the freedom data in the results.

Lines 634-637: For Table 1, I’m confused on all difference letters given. It is too chaos about the sequence of letters listed. Seemingly the a, b, c..... were given randomly! In addition, in the method, 25 eggs per card, replicate 5 times, why 75 eggs were given in the table? In the table, the host eggs of combined species were given, but I can’t find the method description on it!

I also don’t agree to use Female proportion, it is better to change it to percentage of female progeny (%)

Lines 640-644: the same comments to Table 1.

Lines 646-649: the same comments to Table 1.

For Figure 2, change Cold storage period (%) to Cold storage period (d)

Reviewer #2: Dear Authors

Despite a big amount of data was collect and the topic is relavant some weakness was found in your MS.

1) The hypothesis is not very clear.2

2) There isn’t correlation between title, hypothesis and conclusion.

3) Data transformation is not recommended. see the article: “The arcsine is asinine: the analysis of proportions in ecology. Warton and Hui. Ecology. V. 92. 2011.”

4) When you are expressing mass rearing are you are talking about which amount of eggs? 1 Kg per day? This should be considered in order to address a more realistic scale and fit better your finds with the biofactories reality, and also to give for you reader the real idea of what is a mass rearing.

5) In my opinion your data is very useful and interesting, but no great innovation is presented.

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Reviewer #1: No

Reviewer #2: No

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PONE-D-21-01652R1

Cold storage-mediated rearing of Trichogramma evanescens Westwood on eggs of Plodia interpunctella (Hübner) and Galleria mellonella L.

PLOS ONE

Dear Dr. Hasan,

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 Jun 19 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. 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,

Nicolas Desneux

Academic Editor

PLOS ONE

Additional Editor Comments (if provided):

The statistical analyses are not properly presented and therefore their validity is questionable. Because of this the paper can not be accepted for publication in Plos ONE yet. The authors should likely use a GLM analysis instead of doing transformation on datasets. At minimum, authors should present the statistical analyses in a better way (e.g see the comments from reviewer 1).

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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: 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 #1: Yes

Reviewer #2: Yes

**********

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

Reviewer #1: No

Reviewer #2: No

**********

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: Yes

Reviewer #2: 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 qualtiy of updated manuscript has been greatly improved based on the revision suggestions, but there are still serious problems on statistical analyses.

1. lines 184-186: The authors did not give the description in detail. For different trials, seperately describe the statistical methods using ANOVA with multi-factors, what factors have been given for each trial.

2. Table 1: I'm still confused on some difference letters given in table, for example, for the results of adult emergence on 1 day age egg, the data of 87.50 and 91.67 with 'a' and 'ab', respectively. It should give the data: 91.67a, 87.5 ab. Carefully check all the statistical analysis results for Table 1, 2, and 3, then give the right letters for various treatments.

3. Fig. 1: I'm confused on all difference letters given on the bars. It is not clear to note "within eahc egg age, bars followed by the same letters are not significantly different; HSD test at 0.05". Checking Fig. 1, I'm confused on the indication. How you do the comparisions for two species at the same egg age, both togher or seperately? Similarly, the letters on the bars are also given in disorder, check all the statistical results carefully, then give the correct letters for various treatment.

4. Fig. 2: same to Fig. 1. particularly note: for 1 day old egg, at 15 d, why give the letter 'ad' for P. interpunctella?

Reviewer #2: Dear author

Thank you for reply all comments. You presented a very good argumentation, it was good to read it.

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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 #1: No

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.]

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.

Cold storage-mediated rearing of Trichogramma evanescens Westwood on eggs of Plodia interpunctella (Hübner) and Galleria mellonella L.

PONE-D-21-01652R2

Dear Dr. Hasan,

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,

Nicolas Desneux

Academic Editor

PLOS ONE