PONE-D-19-25021

Effect of host fruit, temperature and Wolbachia infection on survival and development of Ceratitis capitata immature stages

PLOS ONE

Dear Dr Papadopoulos,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process.

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Luciano Andrade Moreira, PhD

Academic Editor

PLOS ONE

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1. We noticed you have some minor occurrence of overlapping text with the following previous publication(s), which needs to be addressed:

https://academic.oup.com/aesa/article/95/5/564/28680

In your revision ensure you cite all your sources (including your own works), and quote or rephrase any duplicated text outside the methods section. Further consideration is dependent on these concerns being addressed.

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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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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: No

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 manuscript is technically sound, and the statistical analysis performed appropriately and rigorously. The authors have made all data available.

Overall the information provided in the manuscript is new and interesting. However, the manuscript needs to be edited for grammar. I also found the manuscript too long with the information buried on how Wolbachia infection affected the different fruit fly lines and how the research could be applied. The section on Wolbachia in the introduction should be moved to under the introductory paragraph, and the manuscript also needs to be edited carefully as some sentences did not make sense. For example, lines 62, 82, 83. Also note that it should be Wiedemann NOT Wiedeman.

Reviewer #2: Comments to the Authors:

The authors test the influence of two different host fruits, three different constant temperatures and the population effects of three laboratory lines (Wolbachia-infected and -uninfected) and one semi-wild population on developmental times and survival of larval and pupal stages of Ceratitis capitata. I congratulate the authors for their efforts in this experimental work.

The authors present convincing evidence of a significant influence of each of these variables (and their interaction) on developmental time and survival of C. capitata. The most interesting finding, however, is the influence of Wolbachia on differential survival and performance in the two tested host fruits. The effects of Wolbachia on survival and development of C. capitata in relation to host fruit and temperature is a new finding in this system and brings novelty to the field.

I have minor issues with the English used in this study. I think that some sentences are not clear and a little more elaboration is needed at certain points throughout the manuscript. As there weren’t many writing mistakes, I have taken the time to point some of them out. I still recommend a careful revision of the manuscript. Below, find my comments to the authors.

Minor comments:

>L19: Include the Latin name of species.

>L40: Remove “the” two times. Occurs throughout the manuscript.

e.g. “... expressed lower survival rates and higher developmental times...”

>L44: Not sure I see how “Practical and theoretical implications” are discussed in the discussions. Instead, it appears that implications on the ecology and survival of the fly in nature are discussed.

>L59: Change word “other” to “others”.

e.g. “... some are favourable, while others, are marginal...”

>L60: I think the implications apples have on the over-wintering success of this species in temperate zones is interesting. Perhaps a few more words explaining this could be provided in the introduction.

>L73: Remove parentheses and make meaning clearer. What I think was meant:

“... immatures after being exposed to artificially infested fruit at low storage temperatures for ...”

>L76: I am not sure if the use of the word “biotypes” is appropriate here. In the given literature the word “biotypes” is not mentioned. I would use the word “populations” or “flies” instead. If the word “biotypes” is used, please include an explanation of what is meant by it.

>L78: The term “wildish populations” sounds a little odd. Please make clear that these are populations from the wild that have been reared in laboratory conditions for a certain amount of generations.

>L82: Replace “to an” with “in an”.

e.g. “... this process results in an increased ...”.

>L83: Remove “the”. Replace “on” with “in” and “to a” with “in”.

e.g. “... fitness under artificial rearing conditions and in the same time in reduced performance under different rearing conditions ...”.

I think this sentence, in general, should be double checked.

>L86: Include “as”.

“... factors such as temperature ...”

>L87: Again, the use of “biotypes”. See above (e.g. different medfly lines, populations, etc...).

>L91: Replace “a-proteobacteria” with “α-proteobacteria”.

>L103-109: In this part of the introduction I think it is important to mention how the kind of fitness alterations caused by Wolbachia onto its hosts can be dependent on how long the host is infected with Wolbachia. For example, new or recent infections more commonly result in higher fitness costs than long-standing ones. This will emphasize some differences between natural infections and transinfected lines, where Wolbachia was transferred from a donor to recipient recently. Otherwise, I agree that the relationship between Wolbachia and host represents a dynamic nature not simply explained by any one factor alone and warrants investigation of interactions with abiotic factors.

>L125: Citation “58” is wrongly placed here. Riegler & Stauffer, 2002 did not do laboratory crossings to establish level of CI. They analysed natural populations for infection frequencies. Plus, it is not clear if the authors refer to R. cerasi of C. capitata here.

>L147: Although the term “wildish” is explained in the Materials and methods section, I think it would be good to have a few additional words that explain what is meant by it already in the abstract or introduction. I may be wrong, but the term sounds a little awkward without further description.

>L148-152: Sentence should be re-stated. Was it addressed or wasn’t it addressed?

e.g. “By including Wolbachia-infected lines in our experiments, we addressed whether infection affects ...”

>L158: For reproducibility reasons, it may be worth noting what kind of lights were used (I’d assume this is more important for the adult rearing conditions).

>L161: Italicise the “w” in “wCer2”.

>L171: “... for few months ...” May the authors provide a range or approximation of how many months in more exact terms?

>L176: Closing parentheses missing.

>L194: I think ImageJ may require a citation. Additionally, what was done with these measurements (length and width of pupae)? In the results, it appears that only survival and developmental times were recorded, so it is a little unclear to me what was done with these measurements and how they were incorporated into this study?

> In the supplemental excel file, it is not clear to me how many samples and which samples belong to which of the 20 replicates (L195). This could be clearer. Maybe just by adding a total of how many individuals were finally used.

>L199-204: Although I think the statistical procedures used in this study are sound, I would appreciate to read why they were chosen. Perhaps advantages, disadvantages, or even just references and what exactly was done using them (e.g. tests of significance, interactions, etc...). Also applies to the Wald test, which is only mentioned in the “Results” section (this could be included in the “Materials and methods” section, stating that it was used in the regression analyses to test for significance).

> I recommend adding sub-headers for each section within results and discussions for readability (maybe even materials and methods). E.g. survival rates, performance/developmental duration, etc... Otherwise, the results are presented clearly. The figures and tables are also well made and clear.

>L217: Remove “the”.

“... expressed lower egg-to-pupa survival rates ...”

>L344-348: While I agree that the comparison of wildish flies to the non-infected Benakeio laboratory medfly line supports the idea that reduced genetic variation and an altered microbiome in laboratory lines may result in poorer survival and performance on host fruit, I think it is hard to rule out that the poorer performance of the 88.6 and S10.3 lines is due to the same reasons. Perhaps an additional bottleneck effect might have occurred while trans-infecting these lines and may be worth acknowledging that Wolbachia may not be the sole reason these lines under-perform, especially under higher temperatures where the influence of Wolbachia is supposed to be hampered.

>L438-441: In accordance with the previous comment, if higher temperatures may be detrimental for Wolbachia infections and their influence on hosts, the fact that higher temperatures in this study affected the infected lines most may be indicative of microbiome deficiency or lack of genetic diversity, which the authors include. The authors mention how the microbiome hypothesis could be tested (L421). A genetic/genomic comparison to test the genetic variability hypothesis may be mentioned as well.

>L472-476: These two sentences are not very clear. I’ve missed the take home message of Krainacker et al. Also, next sentence, I think it needs to be re-written for clarity.

> I recommend a thorough inspection of the references section as some inconsistencies and mistakes seem to be present.

Here are just a few examples:

>L611: Reference is missing journal name, volume and page numbers.

>L738: Reference is missing page number.

>L752: Journal name format is different from others.

>L766: Journal name format is different from others.

>L775: Journal name format is different from others.

>L786: Journal name format is different from others.

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Reviewer #1: No

Reviewer #2: Yes: Vid Bakovic

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Effect of host fruit, temperature and Wolbachia infection on survival and development of Ceratitis capitata immature stages

PONE-D-19-25021R1

Dear Dr. Papadopoulos,

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

Within one week, you will receive an e-mail containing information on the amendments required prior to publication. When all required modifications have been addressed, you will receive a formal acceptance letter and your manuscript will proceed to our production department and be scheduled for publication.

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With kind regards,

Luciano Andrade Moreira, 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

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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

**********

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

Reviewer #2: 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

**********

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

**********

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: (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: Vid Bakovic