Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: INVESTIGATING THE POTENTIAL USE OF SENTINEL-1 DATA FOR MONITORING WETLAND WATER LEVEL CHANGES IN CHINA’S MOMOGE NATIONAL NATURE RESERVE
Review round: 1
Reviewer: 2

Output:
Basic reporting: The article titled "Investigating the potential use of Sentinel-1 data for monitoring wetland water level changes in China’s Momoge National Nature Reserve" is well written, well structured, and the figures and tables are significant. The topic is novel and the use of Sentinel-1 adds significant value to the paper.

However, some things need to be addressed before it can be considered for publication:

The references are old. There is no literature review for 2016-2019. There are a lot of studies conducted in the last 2 years that need to be considered.

Please check the following article:
Monthly Analysis of Wetlands Dynamics Using Remote Sensing Data

There is a similarity with the results, you need to compare it with your results.
Experimental design: The methodological part of the study seems logical and meaningful.
Validity of the findings: The results are well structured and supported with meaningful measurements.
Additional comments: Normally I would give this article a minor revision, but the literature leak is a big disadvantage to the manuscript as many studies have been conducted in the last 2 years with Sentinel-1.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: INVESTIGATING THE POTENTIAL USE OF SENTINEL-1 DATA FOR MONITORING WETLAND WATER LEVEL CHANGES IN CHINA’S MOMOGE NATIONAL NATURE RESERVE
Review round: 2
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: no comment


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: MODELLING THE PROGNOSTIC EFFECT OF GLUCOSE AND LIPID PROFILES ON STROKE RECURRENCE IN MALAYSIA: AN EVENT-HISTORY ANALYSIS
Review round: 1
Reviewer: 1

Output:
Basic reporting: The article seems to clearly address most areas implied by basic reporting. While some additional editing is needed for clarity in the English language, and a number of additional clarifications are needed, I think that these should be eminently addressable via revision. Please see my comments to the author below for a line-by-line analysis of the manuscript.
Experimental design: The experimental design appears to be adequate for the question.  Certain elements require additional clarification, however, as listed below in in my line-by line commentary.  Again, these should not be difficult to accommodate.
Validity of the findings: While the overall findings do seem valid, I did note a number of areas in the Results and Discussion, however, where the interpretation given to the data was confusing or questionable.  As above, however, I do not believe this will be excessively difficult to address via appropriate revision.  Please see my line-by-line critique below for specific details.
Additional comments: Thank you for the opportunity to review this study.  This paper uses multivariate regression techniques to investigate key factors associated with recurrent stroke in Malaysia.  While many of these factors have been explored in the western world, little data has yet been published in Malaysia, and this study thus represents an incremental, but important, increase in our knowledge regarding this area of care.  On close review a number of issues were noted that will need to be addressed prior to acceptance, but many of these represent clarifications or enhanced explanations and should thus not be difficult to accommodate.  I did not find any severe methodological flaws.  To aid your team in making the needed corrections, I have structured my critique below according to the article's subheadings.

Overall:

1.Editing is needed for grammar and clarity.

Abstract:

1.Line 40:  I am not certain what is meant by a "contradictory" relationship.  Please clarify.

2.Line 58: While it is indeed possible, the association of triglyceride levels with stroke does not necessarily imply a causative relationship.  Therefore, I think this conclusion needs to be mitigated somewhat.

Introduction:

1.Line 101:  Please delete the word observational.  The same content is implied by your use of the adjacent word exploratory.

2.Lines 107-115:  Please clarify further how the event history analysis approach you describe here represents an improvement on more simple quantifications.  What, specifically, do you expect this approach to yield in terms of greater insight about the relationship between the lab variables tested and stroke risk?

Methods:

1.Overall:  The Introduction implies that event history analysis was used in this study and that this somehow offers more accurate findings.  I did not see this technique mentioned or explained at all, however, in the Methods.   Given its important to your study, please address how your team employed this technique more clearly.

2.Lines 144-145:  It is unclear to me how a time to occurrence variable can be coded as yes/no.  I would expect a scalar time interval to be used instead.  Please clarify.

3.Line 148:  By "censored" do you mean excluded?  Please clarify your use of this word here and throughout the article.

4.Line 149-155: Please provide these numeric concentrations in mg/dl as well as in the current mmol/L notation.  This will enable your results to be more readily interpretable to a wider swath of readers.

Results:

1.Line 173-174: The grammar of this sentence makes it unclear.  Please rephrase.

2.Line 179:  Please clarify what is meant by "25%" significant. What p-value would this correspond to?  The more typical 0.05 p-value does not seem easily relatable to this number and if, in fact, p=0.25 was used at this phase, please state why.

3.Line 179-183:  The sentence is unclear. When you say that you "considered" these, do you mean that you selected likely variables out of the 10 that were significant based on theoretical concerns?  If so, what was the basis for this selection?  Also, why distinguish within the Results between primary and confounding variables. At this point all that you can truly say is that a selection of variables were significant at the designated p-value.  I believe this section should be revised to reflect this, and that the discussion of potential causes versus confounders more properly belongs in the Discussion.

4.Lines 193-204:  This section is a little difficult to follow in terms of what is being presented, especially as concerns the findings of the multiple models.  What I gathered from the statements here is that, for all comers, only total cholesterol had a significant association, but when DM is included as a variable, triglycerides becomes predictive as well.  This is not clear, however, and appears to be contradicted by information within the Discussion.  Please revise for clarity.

Discussion:

1.Lines 214-217:  This is a much clearer statement of the findings of the multivariate regression.  Please revise the Results accordingly.

2.Lines 218-219:  This line suggests that impaired glucose metabolism does predict reoccurrence (albeit weakly), which seems to directly contradict the statement in line 215 that abnormal glucose level does not predict stroke risk.  This may, however, be due to the specific way you are using the terms.  Please clarify.

3.Line 233:  I am confused as to your use of the term "confounding variable" here.  A confounder is typically seen as something which induces unwanted variance that diminishes a study's predictive value.  In this case, however, dyslipidemia seems to be a legitimate predictor, rather than a confounder.  Accordingly, we ask that you consider your use of this term throughout the paper to make sure it conveys what you wish it to.

4.Line 237:  The statement that dyslipidemia and total cholesterol confound seems oversimplified, as the presence of a dyslipidemia would naturally lead to a higher total cholesterol and thus they are in many ways measures of the same thing.  While I appreciate the observation that Malaysian diagnostic bias may indeed render a dyslipidemia diagnosis a partial confounder, the above consideration likely also contributes to the picture.  Overall, more clarity is needed as to how the term "confounder" is being used and why specific variables were classified in this way.

5.Lines 269-277:  While I agree in principle that reducing total cholesterol medically may help with stroke, your actual results only show a degree of association, not a clear causative influence.  For the sake of accuracy and clarity, please consider stating this more clearly either here or in the Limitations section.

In summary, this is a promising article that, while not entirely groundbreaking, will be beneficial to practitioners in Malaysia.  While a number of edits are needed I do not believe they will be difficult to perform, and once these are made I think the article will merit publication.  Thank you again for the opportunity to review your work.  I look forward to reading the next iteration.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: MODELLING THE PROGNOSTIC EFFECT OF GLUCOSE AND LIPID PROFILES ON STROKE RECURRENCE IN MALAYSIA: AN EVENT-HISTORY ANALYSIS
Review round: 1
Reviewer: 2

Output:
Basic reporting: Completely unsuitable . poor English
Experimental design: Design is not adequate : No data is given about risk factors of first ever stroke. No data is given about current or previous therapy (anti hypertensive, anti diabetic etc.., statins etc..)
Validity of the findings: No information about mortality, about atrial fibrrillation and about other important risk factors (for instance anti coagulant therapy)
Additional comments: No additional comments


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: MODELLING THE PROGNOSTIC EFFECT OF GLUCOSE AND LIPID PROFILES ON STROKE RECURRENCE IN MALAYSIA: AN EVENT-HISTORY ANALYSIS
Review round: 2
Reviewer: 1

Output:
Basic reporting: Thank you for the opportunity to review this revised manuscript.  In terms of basic reporting, I found this to be much more readable, and believe that it clearly presents the material in professional scientific English.  There are a few areas that still require copyediting, but this is quite minor.  These are listed below.
1.	Line 136:  The phrases “in males” and “in females” should be outside the parentheses in this sentence.
2.	Lines 174, 175:  The word "triglyceride” in these lines should be revised to “triglyceride level.”
3.	Line 230:  Please remove the “is” after “findings” in this line.
4.	Line 256:  Consider adding a formal “limitations” subheading here.
Experimental design: After reviewing the revised manuscript and reviewer response, I feel that the experimental design is sufficient for the research question and have no further issues or questions.
Validity of the findings: I appreciate the time spent mitigating the strength of the conclusions to better match what a study of this nature can allow in terms of interpretation.  There is, however, one area where such mitigation is still needed.  Line 51 of the abstract suggests that the variables noted as significant within the study contribute to an "increased risk" of stroke.  This, however, cannot be confirmed without prospective study.  I would suggest revising this sentence to state that these variables have "prognostic value" with regard to stroke instead, as this is the interpretation given throughout the body of the article.
Additional comments: I appreciate your attention to the reviewer comments and feel that your article is nearing publishable form.
While a few minor issues exist that require further correction I do not believe that this will prove difficult and, once they are addressed, I anticipate acceptance of your work.  Congratulations on a interesting study.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: THE ENDOPLASMIC RETICULUM-ASSOCIATED MRNA-BINDING PROTEINS ERBP1 AND ERBP2 INTERACT IN BLOODSTREAM-FORM TRYPANOSOMA BRUCEI
Review round: 1
Reviewer: 1

Output:
Basic reporting: In this work, the authors characterised the trypanosome protein ERBP1, which they had previously identified as a potential regulator of mRNA levels in a high-throughput screen. The authors could not confirm the result of this initial screen when they tethered ERBP1 to a reporter RNA, but continued the characterisation of this non-conventional RNA binding protein. They found that the protein is not essential, at least not in the blood stage, but depletion or deletion decreased the growth rate. They show that the protein co-localises with the ER and that it preferentially binds mRNAs encoding ribosomal proteins. They identify one novel binding partner of ERBP1. Overall, this work is of interest to the trypanosome community and most of the data are in sufficient quality and convincing. The manuscript and the figures could have been prepared a little more carefully.

My comments

1) I could not find legends for the Sup Figures. I could therefore not fully judge these figures, in particular S3.

2) The mass spec and RNA seq data need to be submitted to online databases.

3) Please mention the life cycle stage for each chapter/experiment in the text.

4) Once you have a PubMedID, please update the comments on TriTrypDB, in particular, the contradictory results about the tethering experiments.

Figure 1
I don’t see any error bars. I don’t see data of nine experiments. Legend does not correspond to the figure (e.g. –TET is not red). Please correct figure and legend carefully.

Figure 2
The authors use three different nomenclatures, DKO, HKO and ERBP1-/- to name the ERBP1 knock-out cell line, this is confusing, why not stick to just ERBP1-/- (and +/- for the heterozygote). DKO is not even explained. HKO is in particular confusing as it could mean both homo and heterozygous knock out. Doubling time or population doubling time is more accurate then division time. For the procyclic cells, the cell line is labelled HKO in Figure 2E and Figure S2, but this should be heterozygote. Why does SKO grows better than WT?

line 94:
‘Since tetracycline-inducible expression is effected from a very active RNA polymerase I promoter, the result suggests that excess ERBP1-myc is
not toxic. ‘  Without measuring the protein levels, nothing should be concluded as to whether overexpression is toxic or not. A promoter is only one component of many that determines protein levels. Best to omit this sentence.

Figure 5
The localisation of aldolase is not mentioned in the legend (although you mention before in Figure 3). I do not understand the quantifications underneath the bands, looks like percentages, but then, it goes up to 200.  I do not agree to the conclusion from this figure. In B supernatant, ERBP1 looks clearly different from the cytoplasm to me, and not very different from the ER.

Why not load pellet and supernatant of each fractionation next to each other to the same gel and set both bands as 100%? This way, the data would become more comparable. The experiment should be done in triplicates. A graphic can then show averages of either supernatant or pellet with statistics.

Discussion

Line 161: ‘The latter observation is consistent with the exclusion of ribosomal protein mRNAs from trypanosome stress granules. ‘
It would be good to cite.

Ribosomal mRNAs tend to be very small. Is this true for the other associated RNAs too, could size be what matters for ERBP interaction?

Typos etc:

Figure legend for Figure 1: 3x106 (superscript 6)

Line 106: ‘It gave no growth advantage in control cells in which when the marker mRNA had no boxB sequences, ruling out the possibility that ERBP1 expression by itself results in blasticidin  resistance.

Line 118:  ‘To investigate the interactions of ERBP1, we used trypanosomes in which ERBP1 a sequence  encoding a tandem affinity purification (TAP) tag was integrated at the N-terminus, and the other copy of ERBP1 was deleted.’

Line 123: ‘The association with the DED1 helicase may be significant, although we have identified on most other mRNA-related purifications.‘

Typos in Table S2 (legend), purified

Susanne Kramer
Experimental design: all covered in the above
Validity of the findings: all covered in the above
Additional comments: all covered in the above


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: THE ENDOPLASMIC RETICULUM-ASSOCIATED MRNA-BINDING PROTEINS ERBP1 AND ERBP2 INTERACT IN BLOODSTREAM-FORM TRYPANOSOMA BRUCEI
Review round: 1
Reviewer: 2

Output:
Basic reporting: A few places in which the English could be improved.

Abstract: ‘Results from a pull-down’ to Measurements from a puill-down’

Line 33: ‘T brucei grows’ to T brucei proliferates’.
It also proliferates in the tsetse salivary gland.\

Everything else is fine
Experimental design: no comment
Validity of the findings: Line 89:
‘Since some protein always remains after RNAi’
Undoubtedly true but given that the 10 000 fold increase in cell number of the induced RNAi (Figure 1B) how much and where did in come from?

Line 122:
Is there a citation for calmodulin contaminating pull downs?
Additional comments: No additional comments


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: MEDITERRANEAN NEKTON TRAITS: DISTRIBUTION, RELATIONSHIPS AND SIGNIFICANCE FOR MARINE ECOLOGY MONITORING AND MANAGEMENT
Review round: 1
Reviewer: 1

Output:
Basic reporting: The paper includes a sufficient introduction as well as a broad discussion and provides the relevant literature references.
Article structure, tables, figures and raw data conform to scientific claims. The figures are relevant and professional. The submitted paper is self-contained and sound. The results are relevant, but the aims should be complemented and oriented to the title and the discussion respectively (more details see below).
The text is written in clear and unambiguous English, but the word usage should be improved once more by a native speaker.
Experimental design: No comment / not applicable.
Validity of the findings: The study is meaningful, never before there was assessed such a broad selection of traits of Mediterranean fish or other taxa of nekton. I agree with the authors that an investigation of patterns and relationships between these traits is important, especially for the Mediterranean, because the latter is a region that hosts a high biodiversity and has a long history of human influence.
The underlying data have been provided. The authors use simple statistics but it is applicable to the aims of this paper, robust and sound. The conclusions are well stated.
Additional comments: Dear authors,
This paper is well written in large parts, brings novel results and conclusions, and therefore it is eligible for publication in my opinion. However, there are several issues that should be improved.
The most important are following:
(i) Several references cited in the text are lacking in the reference list and vice versa. Please check all references and citations carefully.
(ii) The link to the row data of species and traits given in line 103 does NOT lead to your row data.
(iii) The conclusions (lines 332 – 44) are rather a summary of the discussion than 'conclusions'. This paragraph should be rewritten or omitted.
(iv) Any information about the examined animals (e.g., how many fishes, cephalopods, decapods a.s.o.), would be welcome in the 'Material and Methods'.

(v) As already mentioned above, you stated that the aims of this paper are: (a) to determine whether there are rare or important traits by evaluating the distribution of traits and (b) to detect whether there are undocumented relationships between pairs of biological traits.
But the paper includes much more: a number of findings have significant implications concerning the potential effects of climate change (e.g. through the relationships of the trait of optimal temperature) or habitat loss (derived from the relationships of traits related to tolerance ranges) or the impact of fisheries.
Thus, you should add to your aims that selected 'rare traits' are discussed in relation to climate change and to species' reliance to human impact which also would correspond to the title of the paper. Because you examined many more traits (and even more species) than comparable earlier studies, this part of the discussion is important in my opinion, making the paper especially interesting.

(vi) Does Gambusia affinis play any role in the Mediterranean Sea, or why was it included in your analysis?

You will find some additional remarks in the pdf file.

I hope that these recommendations will be comprehensible and contribute to improve your article.

With kind regards,
Erik Arndt


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: MEDITERRANEAN NEKTON TRAITS: DISTRIBUTION, RELATIONSHIPS AND SIGNIFICANCE FOR MARINE ECOLOGY MONITORING AND MANAGEMENT
Review round: 1
Reviewer: 2

Output:
Basic reporting: In general, the manuscript is well written; the general context is well established and is based on a good knowledge of literature. I greatly appreciate the table 1 which establish the links between traits and the responses of organisms, population, community or ecosystem levels. However, a precise definition of what a trait is would be very useful to avoid confusion and strengthen the scope of the manuscript. Currently, the term “trait” is used in multiple context and the use of different adjectives can generate a misunderstanding. See for instance the use of “biological trait”, “ecological trait”, “functional traits”, “response trait”, “effect trait”…. In this context, Violle et al. (2007) proposed a common framework that can be used such as:
•	Trait: Any morphological, physiological or phenological feature measurable at the individual level, from the cell to the whole-organism level, without reference to the environment or any other level of organization.
•	Functional trait: Any trait which impacts fitness indirectly via its effects on growth, reproduction and survival.
In most literature, functional traits are then divided into 4 types: life history traits, morphological traits, physiological traits and behavioural traits. Ecological traits that refer to environmental preferences are generally excluded even if Costello et al. (2015) in PeerJ 3, e1201 proposed 10 ”priority” biological traits that include both functional and ecological traits.
Personally, I agree with Violle’s definitions which exclude ecological traits. In your paper, many traits are in fact ecological traits which can lead to confusion in interpretations. I therefore suggest that we clearly separate functional traits from ecological traits. Thus, the study of relationships between functional traits alone will make it possible to identify relationships that result from phylogenetic constraints or redundant information between traits (e.g. diet and trophic level), while relationships between functional traits and habitats will identify the influence of the environmental filtering on functional traits. Such a dichotomy should make it easier to discuss the results obtained and to better understand the effects of climate change or fishing on functional traits.
In any case, it is essential to clearly define the terms used in the manuscript.

The figures are relevant but would benefit from more detailed legends; some information is also missing. In Figure 1B, there is no title for the x-axis that corresponds to the rank of species according to the values of the line ranges. In Figure 2, no titles are given to the abscissa and ordinate axes. In parallel with the list of traits in table 2, a definition of each trait should be useful, may be only as supplementary material. In table 3, as the Pearson correlation coefficients matrix is a triangular matrix, only half of the matrix could be provided. It is also misleading to provide non-significant value when the linear regression is not well-adapted. I suggest to replace in the matrix the Pearson correlation coefficients by the regression coefficient.

Raw data (i.e. the database containing the 23 quantitative and categorical traits of fish) is provided as an Excel file that can be downloaded. This database represents a remarkable amount of data compilation work (979 references have been used) that will be extremely useful in the future for all researchers working on biological traits of fish in the Mediterranean Sea. However, I regret that the geographical origin of the data is not explicitly specified as some traits may vary from region to region, or from one habitat to another one. This issue could be also discussed.
Experimental design: The manuscript is a research article which fits well the scope of PeerJ. It is a continuation of an earlier article published by Koutsidi et al (2016) which addressed the same questions on a smaller dataset that only considered traits as categorical variables. The article provides then an analysis of biological traits in Mediterranean marine fish based on a review of a database that provides 23 traits for 235 Mediterranean fish species. It has two major objectives: (1) analyze the distribution of traits in different fish species to identify rare traits and discuss their potential functional role, and (2) study the relationships between traits. The second objective would better defined if the potential confusion in the definition of traits is corrected.
Statistical methods are described in detail and are well-adapted to the data. If there is one weakness, it concerns the ANOVA test. It would be useful to indicate which tests/methods used to verify the normality and the homogeneity of the data prior to the analysis.
Validity of the findings: The discussion is generally based on a complete analysis data. It would benefit from being better organized by distinguishing functional traits from ecological traits and developing some superficially addressed themes. These themes are as follows:
•	The proposal to use functional traits to promote an ecosystem-based analysis of traits by considering all biotic elements (plankton, nekton and benthos) of an ecosystem is relevant is certainly one of the major interests of using traits rather than species identity. It would be desirable to deepen this proposal by suggesting the common traits that could be taken into consideration in these different biotic elements.
•	The authors suggested that the negative relationship between longevity and age at maturity which differs from previous study could be explained by the fact that age at maturity is expressed as the percentage of the life span and not in years. Using the raw data, this hypothesis should be tested.
•	Different authors (e.g. Lavorel & Garnier, 2002; Violle et al., 2007) distinguish ‘response trait’ and ‘effect trait’. I would like the authors to question these notions in the choice of traits to be used to assess the effects of anthropogenic pressures or climate change.
•	To make predictions for future scenarios, the authors argued that anthropogenic stressors may alter trait composition indirectly through the selection of associated traits (see Line 328). If I totally agree with this idea, some authors have also suggested that fishing may induce changes in some traits which are exposed to selective pressures and are heritable (see Edeline, E. et al. (2007) Trait changes in a harvested population are driven by a dynamic tug-of-war between natural and harvest selection. Proc. Natl. Acad. Sci. U. S. A. 104:15799–15804). I would appreciate that this issue on the heritability of some traits is discussed.
•	Traits could be variable among individuals or among populations within a species (see Violle et al., 2017). How can you include this variability in the monitoring program? Do you consider that the trait should be measured from monitoring program only infer from species composition of fish communities?
•	Line 224: Indicate that the reference to Verges et al. (2014) concerns the alien species Siganus recently introduced in the Mediterranean Sea.
Additional comments: The article provides a considerable amount of information on the biological characteristics of Mediterranean fish species. The analysis of the database established during this study provides useful information on the degree of rarity of certain traits and allows discussion of the functional role of rare traits in relation to population size. It also makes it possible to describe the relationships between traits. While this second part of the manuscript highlights expected relationships, it also shows the close links that can exist between functional traits and the environment. Such relationships can help to anticipate the consequences of climate change or anthropogenic pressures on functional diversity. An effort to define the traits and the distinction between functional and ecological traits would go a long way towards ensuring that the work done is in line with the proposed definitions of the traits. It would facilitate the organization of the manuscript and the selection of the main functional traits to be followed in an inventory management context. Some points of the discussion would also benefit from further elaboration.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: MEDITERRANEAN NEKTON TRAITS: DISTRIBUTION, RELATIONSHIPS AND SIGNIFICANCE FOR MARINE ECOLOGY MONITORING AND MANAGEMENT
Review round: 1
Reviewer: 3

Output:
Basic reporting: This study present an impressive database regarding the traits of more than 200 nektonic species from the Mediterranean sea. In addition to the gathering of such a comprehensive dataset that is extremely exciting for future macroecological research on the Mediterranean sea, the authors made a compelling effort to describe the trait distributions and trades-offs at a scale that has rarely been done before. Although the manuscript is well-written, adequately framed and is overall statistically valid, I believe that the major negative point of this study lie in the multiplicity of the tests performed and the lack of a real synthesis of this impressive amount of work (although some notable efforts have been done in this regard). Some of the statistical analyses performed, although mostly valid based on the type of data, results in many figures and tables and could easily be reframed into more synthetic and interpretable analyses. Although it remains perfectly understandable, in its current states the manuscript is hard to follow in some instances, and the key messages sometimes get lost in the amount of information given to the reader. Although I acknowledge the amount of work that some of my major comments may represent, I made several suggestion below that may help reach a more synthetic view of the data. Addressing these several points may I hope add value, understandability and clarity to this work and impressive database, which I feel is necessary in some parts, but in no way these comments should impede the publication of this work.
Experimental design: no comment
Validity of the findings: no comment
Additional comments: --------------------
Major comments
--------------------

------------1---------

The major drawback of this study is the consideration of only pairwise relationships, which results in many graphs and tests. Although I do not dismiss the value of these tests, they are hard to synthesize and do not considered the multicollinearity inherent to multivariate data. An easy way to summarise the groups of traits that covary and account for the multiple relationships existing among traits is to resort to multivariate analyses. Such multivariate analyses could be complemented by the existing tests, that are fully complementary to the approach I suggest below, but theses bivariate tests should only be for me an additional information, the core information being the description of the trait space through multivariate analyses.
Given the mixed type of the traits considered here, Hillsmith ordination (Hill and Smith, 1976) appears to be the most appropriate tool. The latter can be performed easily in R using the dudi.hillsmith function of the `ade4` package. See for e.g. Teichert et al. 2017

I think using such a multivariate approach could also broaden the scope of this manuscript in several ways. The authors have focused in several instances on the concept of rarity and such an approach would allow an easy identification of rare traits and of the species bearing them (the keystone species concept discussed in l. 63 and l.218 but that cannot be found anywhere in the results). In addition, it could allow to identify parts of the trait space with high redundancy, a concept that has been eluded in this manuscript and that should be more emphasized as it is as important as rare traits in terms of conservation and would allow a more comprehensive depiction of the Mediterranean nektonic trait space. See for e.g. Diaz et al. 2016 for a nice example on how to use multivariate approach to make a comprehensive description of trait spaces.

In addition, to go further on the idea of keystone species that I find particularly relevant and in the aim of reducing the complexity of the data and synthesizing information, functional groups could be delineated using clustering techniques as performed for example for Mediterranean copepods (Benedetti et al. 2016). It is fully complementary to the description of the trait space with ordination and the functional groups delineated could for example be represented on the ordination (see for e.g. Figure 4 of Benedetti et al. 2018).

Although the clustering of functional groups is more optional, I feel the inclusion of
an ordination (with something like the Figure 2 of Diaz et al. 2016) really necessary for synthesizing the results, simplifying the discussion regarding rare traits and traits trade-off, and developing more on the concept of keystone species and on the degree of redundancy of the Mediterranean nekton.

----------2------------

The choice of the 235 species should be more thoroughly discussed: why these 235 species and how representative is it of the diversity of nektonic species of the Mediterranean Sea? How much of the trait distribution you show here is actually relevant at the Mediterranean scale and how much of it is due to the species included and only described the database rather the Mediterranean nekton? On a similar note, does the database comprises of rare species? Is there commercial ones? I believe that answering these different questions with additional details on the pool of species included in the database would promote the reusing of this extensive dataset and would clarify the relevance of your results in terms of the ecology of the Mediterranean nekton.

---------3-------------

Although I feel several of the figures should be relegated to the Supplementary material with the inclusion of a multivariate approach (potentially Fig 3 and 4), I do have several comments on the understandability and readability of the figures and tables.
Fig 1 and 2 are fully relevant here and extremely interesting but I have some doubts about the representation chosen for continuous traits. Density distribution should be used instead of histograms and the threshold discussed in the text could then be highlighted through colors or shading (for e.g r - Shading a kernel density plot between two points. - Stack Overflow : https://stackoverflow.com/questions/3494593/shading-a-kernel-density-plot-between-two-points)

For Fig 1B, the x-axis should be named and it should be mentioned in the legend that individuals are ranked according to the mean of the range (if that is what is done). Also, there seems to be different shades of grey according to the bars. I am not sure what is the meaning of these.

I would prefer the panels of Fig 2 to be all on the same y-scale and to range from 0 to 100 to make the different dominance patterns clearer.

In Fig 3, in table 3 and in the text, the distinction made between linear and non-linear relationships are somehow obscured by the presence of log-transformed variables. I believe a clarification is needed here since a linear relationship between two log-transformed variables means cannot be called “a linear relationship between these two variables” as it is rather log-linear. Therefore, not only three pairs of variables show non-linear relationships (as said l. 185) but most of them do strictly speaking.

Also, if something like Table 3 and 4 was to be kept along with the multivariate analysis (for e.g. to support the significance of the relationships observed in the ordination), I would rather see one lm or glm per traits with all other traits (and potential polynomes of these traits) as explanatory variables and a table reporting the standardized slopes of each explanatory variables with the R2 and significance of the full model. This could allow to gather Table 3 and 4 into a single analysis where all variables (quantitative and qualitative) would be included. A selection of variables (through AIC for example; stepAIC function) would allow to reduce each model, and then the coefficients of the selected variables only would be reported into a single table reporting the best model for each traits (models that would contain potentially both continuous and categoric variables, as the coefficients of both are perfectly interpretable). See :

Schielzeth, H. (2010). Simple means to improve the interpretability of regression coefficients.Methods in Ecology and Evolution,1(2), 103-113.

Zuur, A. F., & Ieno, E. N. (2016). A protocol for conducting and presenting results of regression‐type analyses.Methods in Ecology and Evolution,7(6), 636-645.

Fig. 4 should be reorganized, either to group modalities belonging to the same trait together, or to group modalities with similar co-occurence patterns together.

---------4--------------

The choice of forcing each species to belong only to one category of a trait instead of allowing them to belong to more than one category using fuzzy coding, which allows accounting for some of the plasticity the species (Chevenet et al., 1994), should be at least discussed. Although fuzzy coding may be dependent upon expert knowledge when giving relative scores to the affinity of species to each modality of a trait, it seems a more realistic view of the plasticity of the species. The coding approach has a deep influence on the subsequent patterns described. Therefore, it should be made clear to the readers in the Material and Method section the type of coding that has been used and I believe that a quick discussion (that could potentially be a perspective of this work) on the matter would add value to the manuscript.

--------------------
Minor comments
-------------------

Thank you for providing the data, this will make a great contribution to future research. However, blank should preferentially be coded with “NA”. Also, if all data seem available, the range for “range type” traits are not provided (only the mean is), which impedes the reproducibility of Fig 1B and would also be an interesting information to make available in this database for future studies.

l.40. Functional traits “impact/influence” directly or indirectly the fitness of organisms but do not always increase it. Please change the “increase” accordingly to match with Violle’s definition of functional traits

Table 1 could be reorganized to put traits that describe similar features together (e.g. Traits 8/9 with traits 13/14/15; or trait 19 with trait 12; or traits 17/18 with trait 20).

l.47 to 64. The framing of the study is well done but I feel that mentioning references on key works that have described trait trades-off for other taxonomic groups and how they have allowed a more comprehensive and mechanistic understanding of these groups may make an even better case for this study (e.g. Litchman et al. 2013)

l.63. KeyStone instead of keynote

l.88. I would change “important” for “dominant” traits

l. 145-155 and l.236-241 : These results on the distribution of traits could be compared to other existing works to better put them in perspective. For e.g., although the categories included are not fully similar, I feel that discussing the similarities and differences in the proportion you find with those from the North Altantic and Pacific (See for e.g. Supplementary Figure 2 of Beukhof et al. 2019) may be worthwhile. Especially, herbivorous species are low while benthivorous species dominates in both databases. On the other hand, you did not include a “generalist” modality (which I think is a good thing) but this should be mentioned and discussed as it changes the relative proportion of other traits (especially piscivorous species apparently). Such comparison would allow to extend the generality of your findings and open-up towards some potential general trends regarding the structure of nekton trait space.

l.149-150 Can you also indicate the mean, median or mode of the distributions, as well as potentially the range of some variables when relevant

l.157 I am not sure of what you mean by “angle” and did not understand this sentence. Could you please clarify your thoughts ?

l.243 : weighting may be done using species abundance I agree, but may/should also be weighted by biomass (especially when looking at the role of traits on ecosystem rates and functioning)

l.253 Another way (and potentially more conclusive one) to address and confirm the non-random distribution of traits among species would be to assess whether or not these >200 nektonic species are distributed randomly in trait space using a randomization approach such as the ones performed in Diaz et al. 2016. However, I acknowledge that this may not fully fall within the scope of this study, but it may nonetheless be an interesting open-up/perspective.

l.298 In my opinion, many of the traits you included are rather « soft » (each traits are proxy of one or several functions rather than direct measures of these functions). Although the distinction between hard and soft is rather subjective, I feel many of these traits do not fit in the definition of hard traits given by Violle et al. 2007 (Oikos) or Nock et al. 2016 (Functional traits). Some of them such as temperature/depth range would better fall in the category of “ecological traits” as defined by Beauchard et al. 2017 (and see the potential limitations discussed in this paper for the use of these traits). Therefore, I would remove this term that is for me more than questionable here.

l.306-308. The distinction between effect and response traits may be relevant to formalise the distinction you are making here (see Suding et al. 2008)

l.326-329 : Suggested ref for the differential response to stress among fish functional groups and the need to understand traits to understand their responses : Henderson, C. J., Gilby, B. L., Schlacher, T. A., Connolly, R. M., Sheaves, M. , Maxwell, P. S., Flint, N. , Borland, H. P., Martin, T. S., Gorissen, B. and Olds, A. D. (2019), Landscape transformation alters functional diversity in coastal seascapes. Ecography. doi:[10.1111/ecog.04504](https://doi.org/10.1111/ecog.04504)

l.326-329 : Suggested ref for the need to incorporate trait and life-history knowledge of nektonic species to understand their dynamics : Marquez, J. F., Lee, A. M., Aanes, S. , Engen, S. , Herfindal, I. , Salthaug, A. and Sæther, B. (2019), Spatial scaling of population synchrony in marine fish depends on their life history. Ecol Lett, 22: 1787-1796. doi:10.1111/ele.13360

Table 5, penultimate line : temperature instead of temp/ture

----------------
Suggested ref
----------------

Beauchard, O., Veríssimo, H., Queirós, A. M., & Herman, P. M. J. (2017). The use of multiple biological traits in marine community ecology and its potential in ecological indicator development.Ecological Indicators,76, 81-96.

Fabio Benedetti, Stéphane Gasparini, Sakina-Dorothée Ayata, Identifying copepod functional groups from species functional traits,Journal of Plankton Research, Volume 38, Issue 1, January/February 2016, Pages 159–166,https://doi.org/10.1093/plankt/fbv096

Benedetti, F,Vogt, M,Righetti, D,Guilhaumon, F,Ayata, S‐D.Do functional groups of planktonic copepods differ in their ecological niches?.J Biogeogr.2018;45:604–616.https://doi.org/10.1111/jbi.13166

Chevenet, F., Dolédec, S., & Chessel, D. (1994). A fuzzy coding approach for the analysis of long‐term ecological data. Freshwater Biology, 31(3), 295–309. https://doi.org/10.1111/j.1365‐2427.1994.tb01742.x

Díaz, S., Kattge, J., Cornelissen, J. H., Wright, I. J., Lavorel, S., Dray, S., … & Garnier, E. (2016). The global spectrum of plant form and function._Nature_,_529_(7585), 167. The global spectrum of plant form and function | Nature

Hill, M. O. and Smith, A. J. (1976), PRINCIPAL COMPONENT ANALYSIS OF TAXONOMIC DATA WITH MULTI‐STATE DISCRETE CHARACTERS. TAXON, 25: 249-255. doi:10.2307/1219449

Litchman, E., Ohman, M. D., & Kiørboe, T. (2013). Trait-based approaches to zooplankton communities.Journal of Plankton Research,35(3), 473-484.

Nock, C. A., Vogt, R. J., & Beisner, B. E. (2001). Functional traits.eLS, 1-8.

SUDING, K. N., LAVOREL, S. , CHAPIN, F. S., CORNELISSEN, J. H., DÍAZ, S. , GARNIER, E. , GOLDBERG, D. , HOOPER, D. U., JACKSON, S. T. and NAVAS, M. (2008), Scaling environmental change through the community‐level: a trait‐based response‐and‐effect framework for plants. Global Change Biology, 14: 1125-1140. doi:10.1111/j.1365-2486.2008.01557.x

Teichert, N., Pasquaud, S., Borja, A., Chust, G., Uriarte, A., & Lepage, M. (2017). Living under stressful conditions: Fish life history strategies across environmental gradients in estuaries.Estuarine, Coastal and Shelf Science,188, 18-26. Redirecting

Supplementary material of Beukhof, Esther; Dencker, Tim S; Palomares, M L D;Maureaud, Aurore(2019):A trait collection of marine fish species from North Atlantic and Northeast Pacific continental shelf seas._PANGAEA_,(https://doi.org/10.1594/PANGAEA.900866)


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: CHARACTERIZATION OF A MN-SOD FROM THE DESERT BEETLE MICRODERA PUNCTIPENNIS AND ITS INCREASED RESISTANCE TO COLD STRESS IN E. COLI CELLS
Review round: 1
Reviewer: 1

Output:
Basic reporting: Manuscript for the most part written in clear and professional manner. However, there are some sentences that would need revision. It is suggested that the authors proofread the manuscript for some grammatical errors.
Experimental design: The identification and characterization of Mn-SOD from Desert Beetle and testing its cold-resistance property in E. coli is a novel study.
Major comments:
1)	Methods section needs to be more detailed. Specifically, details on statistical analysis in methods only mention Tukey’s post-hoc test alone. It does not mention one-way ANOVA as its primary statistical test run by authors for identifying significant changes in their treatment data. Moreover, some of the experiments were analyzed by paired Student’s t-test which needs to be mentioned in the methods.
2)	There is some detail on how relative electrical conductivity assay was performed and results analyzed in the supplementary file However, authors should explain the complete procedure in the methods section.
3)	Though the overexpression data in E. coli is well put together and supports the hypothesis strongly, it would be great if authors could in near future show protection from cold stress by Mn-SOD via knock-down or knock-out models for Desert Beetle.
Validity of the findings: No comments
Additional comments: No additional comments


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: CHARACTERIZATION OF A MN-SOD FROM THE DESERT BEETLE MICRODERA PUNCTIPENNIS AND ITS INCREASED RESISTANCE TO COLD STRESS IN E. COLI CELLS
Review round: 1
Reviewer: 2

Output:
Basic reporting: This paper is a good follow-up of their previous publication based another two enzymes called “Cu/Zn-SODs”. Overall the paper submitted in PeerJ is well-written with sufficient intro about the background of the enzyme Mn-SOD, good results and informative discussion. The abstract is also clear and can summarize what they have found in this paper.
The cold-induced upregulation of MpmMn-SOD is interesting and I think the author has provided some results (figures and table, etc.) to support their claim, with appropriate references. However, I still have some concerns about the data shown in this paper and the consistency of the enzyme activity between the beetle and the E-coil. I will describe these concerns in the following parts.
In general, I think I recommend this paper to be published in PeerJ if the authors can address all my concerns.
Experimental design: The experiment design in this paper is similar to the Cu/Zn-SODs paper, except that the model enzyme used here is the MpmMn-SOD. Therefore, I think the research method described in this paper is systematic and solid-built. However, I still have some concerns about the experiment mentioned in this paper.
1. As the authors mentioned in the intro part (line 77), the average temperature of the soil the beetle would confront in Jan are -12 C to -5 C.  However, the experimental temperature in this paper for beetle individuals was performed at 4 C. I am more concerned that the temperature difference will affect some of the data shown in the paper. I am also curious about the reasons that the authors wouldn’t perform these experiments in -5 C. Maybe the authors should add some sentences in the experiment part to address this. I did find out that the E-coil experiment was performed at -4 C. I think this temperature is much closer to the real temperature the enzyme will suffer.
2. The mRNA experiment shown in Fig. 4 is interesting but confused. In A it shows that the hindgut and fat body rank the first and second tissues that have more MpmMn-SOD expression. However, in B it only shows the expression of the overall mRNA in the beetle. In my opinion, B needs to show the different expressions in all the tissue listed in A, at least show the data in hindgut and fat body would be better. Also the data is compared to the head at 0h, which would give you the highest fold-change as the head has the lowest amount of MpmMn-SOD. But why you don’t compare it to the other 4 tissues listed in A?
3. Again, in figure 4B, at 11h the mRNA level seems to go back and it is very interesting. It looks like another round of cold-defense is going on? I recommend the authors could briefly explain this in results and discussion.
4. The SDS-PAGE experiment in Fig.5 A, there is a clear and strong up-regulation band in lane 2, located in ~20 KDa, what is this? It seems that the band disappeared in the lane 4. If lane 2 and lane 4 are both induced samples and the expression of pET32a-mMn-SOD wouldn’t affect the ~20KDa protein, should it be also on the lane 4, same position? Can you explain this?
5. Since the data in Fig.4 showed the experiment till 11 h (11 h shows a growing again and is interesting), it would be better that the same time data points (8h, 9h, 11h?) are shown and compared in Fig.6. Then you can compare the activity of the enzyme both in beetle and E-coil.
6. The Oxford-cup experiment in Fig. 7 is also interesting. Although the control showed a bigger circle compared to the MpmMn-SOD one at same H2O2 concentration, the circles in the control experiments are very dim and absent of a clear, sharp circle boundary. Why this happened? Can you explain?
Validity of the findings: Overall, I think the paper have shown some interesting findings. I already mentioned lots of comments in the previous section.
Additional comments: I think this paper is a good fit to PeerJ if the authors can address all my concerns lised above. Those comments are my main concerns about the paper.
Again, I have some minor revision, typo stuff for the authors, too.
1. The term BL21 in the abstact is very confused to me. I don't think that people would know it is a E-coil if they are not cell biologists. Maybe add it after "An E. coli system" in line 24, such as "An E. coli system (BL21) was applied ...."
2. Line 45, "reactive oxygen species" should be "ROS" as you mentioned the acronym before.
3. Line 53, "Superoxide dismutases" should be "Superoxide dismutases (SODs)", as you used SOD below.
4. Line 62, "MnSOD" should be "Mn-SOD".
5. Line 144, "MpmMnSOD" should be "MpmMn-SOD".
6. Sentence from Lines 301 to 302, please rewrite it.
7. Lines 403 and 411, two "futher"s seem weird. Could change the second futher to some other words, such as "last but not least"?
8. Line 542, the reference is wrong. The doi is same as the reference in line 538. Also I couldn't find the reference.
9. In SI material all the "datas" should be "data". "Data" is used as a plural noun in technical English, when the singular is datum.
10. Figure 5 labels A and B in figure are missing.

Overall, I recommend a major revision of the paper.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: CHARACTERIZATION OF A MN-SOD FROM THE DESERT BEETLE MICRODERA PUNCTIPENNIS AND ITS INCREASED RESISTANCE TO COLD STRESS IN E. COLI CELLS
Review round: 1
Reviewer: 3

Output:
Basic reporting: 1. The paper requires corrections of a few errors in the text e.g. line 382 where the time required to reach 125-fold is missing.
2. The description and the y-axis title differ in figure 6B.  Is it O2- content or O2- content reduction?
3. I would encourage the authors to include a section with commonly used acronyms.
4. Do insects have any other antioxidant enzymes?  I believe it would enhance the knowledge of the reader if you were to mention in the introduction, others that might participate in the same process.
5. 2.	Figure 2: Can you please add to your description, what the colors cyan, pink and black represent?
6. 3.	What was the purpose of the analysis behind figure 3?  Is there anything you can infer based on the classification about the behavior of the protein?  Can any similarities be drawn with the enzyme version expressed by T. molitor?  While you mention it in a few lines (231-235) in the results section, a bigger discussion about the different clades of enzymes in the introduction or results would help.
7. 5.	Figure 6: Why does the SOD activity increase not correlate with a drop in O2- levels?  What do you hypothesize as the reason for increase in O2- levels at 6h?
8. Does the over-expressed enzyme in BL21 bacteria localize to the mitochondria and if it does not, would that affect its effective activity?
Further, I encourage the authors to have the manuscript proof read.
Experimental design: No comment
Validity of the findings: In the experiments leading to Figure 6, it is mentioned that OD 595 readings were taken at the end of the assay.  Would you mind sharing this as well?  I would like to see how viability is affected by prolonged treatment to low temperatures and how this might affect the data.
Additional comments: No comment.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: CHARACTERIZATION OF A MN-SOD FROM THE DESERT BEETLE MICRODERA PUNCTIPENNIS AND ITS INCREASED RESISTANCE TO COLD STRESS IN E. COLI CELLS
Review round: 2
Reviewer: 1

Output:
Basic reporting: A. The manuscript still needs proofreading. There are minor language errors throughout the manuscript that may create confusion. Some of the changes have been made to the "tracked changes" document and "rebuttal" which will be forwarded to the editor.

B. The authors have not added to discussion that testing the hypothesis with knocking down the enzyme in Desert Beetle is missing from current manuscript or is a weakness that they would address in future.

C. "To let the results be clearer to read Fig.6 was modified and supplied with the measured values of SOD activity (Fig.8A) and O2•- contents (Fig.8B), respectively (because OD595 readings were supplemented as in Fig6 in the revised version, the original Fig.6 was remarked as Fig.7, and so on for the following figures); The corresponding relative differences were given in Fig. 8C and Fig.8D, respectively".

Whatever the authors are trying to convey in this part of rebuttal is not clear at all. Kindly revise.
Experimental design: A. The authors have perhaps not understood the second reviewer’s comments about the mRNA experiment. In figure 4B, it is still unclear what the control at 0h is. In the correctly performed experiment, it should be mRNA level of the same tissue at 0h. Secondly, the labeling on top of bars (c, b, cd etc.) have not been explained in the legend. Please rectify these issues.

B. Please specify how the 4 concentrations for hydrogen peroxide for Oxford-cup experiment chosen.
Validity of the findings: No comment.
Additional comments: No additional comments


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: CHARACTERIZATION OF A MN-SOD FROM THE DESERT BEETLE MICRODERA PUNCTIPENNIS AND ITS INCREASED RESISTANCE TO COLD STRESS IN E. COLI CELLS
Review round: 2
Reviewer: 2

Output:
Basic reporting: The authors have addressed all my concerns very well. I think right now the revised manuscript meets all the criteria that PeerJ requires. I recommend the acceptance of the manuscript.
Experimental design: no comment
Validity of the findings: no comment
Additional comments: I only found some small typos in all the figure captions, which are located after the main pdf manuscript. Some sentences are repetitive. Please revise them in the final submitted version.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: CHARACTERIZATION OF A MN-SOD FROM THE DESERT BEETLE MICRODERA PUNCTIPENNIS AND ITS INCREASED RESISTANCE TO COLD STRESS IN E. COLI CELLS
Review round: 2
Reviewer: 3

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The authors have addressed all my comments satisfactorily.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: IMPLICIT MOTOR IMAGERY PERFORMANCE IS IMPAIRED IN PEOPLE WITH CHRONIC, BUT NOT ACUTE, NECK PAIN
Review round: 1
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The authors reported from an on-line study accuracy and reaction time of 1404 individuals with and without neck pain for two (neck and hand) laterality judgement tasks. They performed different analyses when grouping participants under 3 status (neck pain status, pain location and head rotation-evoked pain). Overall, individuals with chronic neck pain were less accurate but not slower than individuals without pain (main effect of neck pain status). When grouped with pain location and head rotation-evoked, the results are more disparate. The authors concluded that individuals with chronic neck pain have disrupted proprioceptive representations, without an influence of pain location and movement-evoked pain.
Please find below general concerns and minor comments:

Abstract
- The results when individuals were grouped on pain location status are confusing. The authors should only present main findings that help understanding the general message. Also, the authors should clearly identify the different analyses depending on the grouping status to help the reader to better understand the results (for example, after ‘Of 1404 participants, 105 reported acute neck pain and 161 reported chronic neck pain’, they could write ‘When grouped on neck pain status, people with […].  When grouped on pain location, […]’ or something equivalent).
- l.50: When saying ‘People were faster and more accurate’, does this mean that a test was performed for accuracy and another for RTs? However, only one p-value was reported (p<0.001).

Introduction
- The authors could provide further information to understand the relevance of having Aim 2 and Aim 3, especially when results of aim 2 and 3 are (almost) similar?
- l.137-139: the hypotheses are not in line with the Aims as the authors did not talk about comparison with hand left/right judgement task (l.133-137).

Method
- l.198-203: while the comparison between neck and hand laterality judgement tasks is relevant, how did the authors deal with the fatigue, as mentioned l.201 (the hand tasks being the fifth block)?
- l.210-211: ‘if the response time for eight consecutive images […] data from those images were excluded’. Why eight consecutive images, and not each time the trial lasts 5 seconds?

Statistical analyses
- It said line l.235 that an ANOVA was performed to assess for a possible speed-accuracy trade-off. What was the variable tested?

Results
- l.329-340: the main message for these results is not clearly presented. The authors should try to reformulate this paragraph, only presenting the significant results of the interaction Direction of Image Head Rotation x Side of Pain interaction.
- l.361-362 ‘There were no clear location-specific effects on performance – only differences based on pain location were found’. This seems contradictory.
Why was the analysis on hand laterality judgement task only presented for Aim1?

Discussion
- Did the authors ever consider the intensity of neck pain as a possible co-variate to explain the results? This could be mentioned in the limitation section of the discussion.

Minor comments
- l.226-227: were accuracy responses normally distributed after log transformation?


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: IMPLICIT MOTOR IMAGERY PERFORMANCE IS IMPAIRED IN PEOPLE WITH CHRONIC, BUT NOT ACUTE, NECK PAIN
Review round: 1
Reviewer: 2

Output:
Basic reporting: The ms is generally well-written and referenced. A few minor comments:
Line 518: What specific theories?

The authors state “motor efferent copies (i.e., a copy of the motor command sent) are created and used to determine if the movement output occurred as planned. Theories with which I am familiar typically assume that efferent copy underlies the “forward model” and allow prediction whereas sensory feedback permits one to determine if the movement occurred as planned. Please clarify this.

In line 303 the authors refer to performance at “identifying an image of a neck” rather than identifying the direction of head/neck turn; this should be corrected.
Experimental design: The design is generally adequate but several points should be addressed.

The absence of information about pain severity is striking and a significant limitation. It is certainly possible that pain severity rather than chronicity or “location” of pain is the primary determinant of performance. Pain severity ratings are always subjective and potentially confounded by other factors, but it is not clear that the ratings would be more unreliable if provided on-line than in an interview setting.

Why were 4 blocks of data collected but only the second and fifth blocks reported in the manuscript? If fatigue was anticipated, why were the 3 and 4th blocks included? Was there evidence from behavioral measures (e.g., RT) to suggest that fatigue was an issue? If so, what are the implications for the hand laterality data? Were the results different if all of the data were included?

How much data was eliminated because of timing out or too short responses?
Validity of the findings: The findings appear solid and the analyses generally well motivated.

The major weakness of the manuscript in my view is the discussion. The authors conclude that the data “support the presence of impaired neural proprioceptive representation of the neck in people with chronic neck pain”. This conclusion is not adequately supported. Nor is the self-referential conclusion that deficits in accuracy are more consistent with proprioceptive deficit and deficits in RT are more consistent with unequal weighting. Although one wouldn’t expect an exposition similar to that presented in the reviews cited, this claim needs to be developed; as the discussion is long and at some points rambling (e.g., speculation about the absence of effects of side of pain), there is plenty of space to present evidence relevant to this point.

From the early work of Parsons to the present, many investigators have drawn conclusions based on differences in RT as a function of stimulus type (e.g., the degree of angular deviation of the stimulus). As these knowledgeable authors well, know, with normal subjects there is a very characteristic RT pattern for hand and foot identification tasks, presumably reflecting the time taken to mentally rotate the body part to match the pictured stimulus. RTs and accuracy information should be provided for each type of stimulus (e.g., 60° right head turning; left hand palm up at 270°) rather than just summary measures collapsed across direction and magnitude of head turning.  Such an analysis may be useful in addressing the question of a relationship between neck position and location of pain, for example.
Additional comments: In general, I thought this to be an interesting and generally well-done manuscript.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: IMPLICIT MOTOR IMAGERY PERFORMANCE IS IMPAIRED IN PEOPLE WITH CHRONIC, BUT NOT ACUTE, NECK PAIN
Review round: 2
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The authors responded point-by-point to my comments.
I have no further remark.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: PHYLOGENY OF LIBELLULIDAE (ODONATA: ANISOPTERA): COMPARISON OF MOLECULAR AND MORPHOLOGY-BASED PHYLOGENIES BASED ON WING MORPHOLOGY AND MIGRATION
Review round: 1
Reviewer: 1

Output:
Basic reporting: It was my pleasure the review this article and I congratulated the authors on an interesting piece of work. The manuscript is well written and clearly structured. The use of English seems professional throughout, with a few exceptions (see below).
The background provided helped me, as a non-entomologist, understand both the research objectives and the results obtained. The raw data are shared as supplemental information.
Altogether, the article is self-contained and convincingly conveys its message.

I have a few linguistic issues to consider:

line 32: Paup misspelled
lines 71-74: reading these two sentences, I have difficulties understanding the difference between the two views. Please rephrase.
lines 81-84: what do you mean by "population evolution law" and "systematic generation relationship"? Please explain or use other expressions here.
line 92: delete "study"
lines 102-103: what is the "law of genetic difference"? I am not familiar with this expression.
line 109: is "coevolution" the right word to use here? I think you need to reformulate your ideas here.
line 129: check spacing at brackets
line 130: delete “its”
line 139: verb missing?
lines 139-140: “Dragonfly” is not a species at all. Please rephrase, either by mentioning the actual species you refer to or by referring to dragonflies as a clade (specify the relevant taxonomic rank).
Caption of Fig. 1: check spacing
line 299: “hindwing” instead of “findwing”
line 316: “dragonfly species” instead of “species dragonflies”
lines 385-386: “due to homoplasy” instead of “due to the trait homoplasy”
line 392: replace “so” by a comma
Caption of Fig. 7: “The clustering of the forewing…” instead of “he clustering of the forewing…”
Acknowledgments: this section includes a funding statement. As far as I understand, this is not the right place for funding statements according to journal guidelines.
Experimental design: The research presented seems novel to me and falls within the scope of the journal. It addresses an unambiguously defined aim that seems both relevant and meaningful to me. The match or mismatch between molecular phylogenies and morphological characters is a highly interesting but still poorly understood topic. The more rigorously performed studies, such as the present one, the better our knowledge.
For some of the methods described, I lack the necessary expertise to judge the validity of the details provided in full extent. The phylogenetic and morphometric approaches, however, seem sound to me.
Validity of the findings: The findings presented seem valid to me, well-presented and supported by the data provided. I cannot assess the statistical soundness of all data so it would be useful to let someone with the relevant expertise review this aspect specifically.
The conclusions presented refer to the research questions raised and seem perfectly sound to me.
Additional comments: Good job! Looking forward to seeing this published.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: PHYLOGENY OF LIBELLULIDAE (ODONATA: ANISOPTERA): COMPARISON OF MOLECULAR AND MORPHOLOGY-BASED PHYLOGENIES BASED ON WING MORPHOLOGY AND MIGRATION
Review round: 1
Reviewer: 2

Output:
Basic reporting: Are the interspecific relationships established based on genetic information consistent with the traditional classification of Libellulidae dragonflies which is based on the wing shape?
This is an interesting question, which the authors, aimed to answer that by combining geometric morphometrics method, principal component analysis (PCA), cluster analysis (UPGMA), etc.
The manuscript is well written, introduction is well motivated, methods are suitable, and results are well presented.
Experimental design: The only drawback is small sample size (the authors focused on only 10 species from a single family), which to some extent narrows the scope of the manuscript and raises the question whether the results can be generalized to other species and families or not. However, it still facilitates more comprehensive future studies.
Validity of the findings: All criteria were met.
Additional comments: I would like to suggest the acceptance of the paper with following minor comments.
1.	Something that I was missing is the images of the wings analyzed in this study in comparable size, which can be added as suppl mat.
2.	Line 116
this sentence would benefit from a more relevant citation.
3.	Line 139
needs revision.
4.	Line 279
Based on
5.	Line 298
for a non-specialist, it would be nice to denote “apical”, “radial”, etc. in a wing image.
6.	Line 299 and line 300 “dominated by the wing nodus”
What does this exactly mean?
7.	Line 299 “findwing”
8.	Line 301 “is the basis of the whole wing structure”
It is a vague expression.
9.	Line 302 “wing nodus is the main load-bearing”
Rajabi et al, in their paper, indicated that the nodus plays a key role in wing deformation and likely subjected to extensive bending and torsion in flight.
10.	Line 313-315
These sentences require citation.

Hamed Rajabi, D. Sc. D. Eng.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: GYNODIOECY IN THE COMMON SPINDLE TREE (EUONYMUS EUROPAEUS L.) INVOLVES DIFFERENCES IN THE ASYMMETRY OF COROLLA SHAPES BETWEEN SEXUALLY DIFFERENTIATED FLOWERS
Review round: 1
Reviewer: 1

Output:
Basic reporting: This manuscript entitled « Gynodiecy in the common spindle tree (Euonymus europaeus L.) involves differences in the asymmetry of corolla shapes between sexually differentiated flowers » by J. Neustupa submitted to PeeRJ is an exemplary interdisciplinary study integrating botany and mathematics.

Figures are relevant, of high quality, well labelled and described.
Figure 1:  I could not see the scale bar on the images.

The English language is clear and professional throughout the manuscript, and it was a pleasure to read this article.
Experimental design: The geometric morphometric methods applied here were published recently, and the conclusions that are drawn are convincing, interesting, and open the way to a whole set of studies of floral symmetry using this novel approach.

The research question is well defined, and the experimental design is perfectly adapted to answer the question.
Validity of the findings: The findings are well supported by the data; they are original and well interpreted.
Additional comments: The species that is investigated is an emblematic species in evolutionary biology and belongs to the set of plant taxa which has been studied by Darwin. This will draw the attention of many readers, may they be botanists interested in the evolution of flower shapes, morphometricians keen on investigating organismal symmetry, or ecologists curious of the relations between flower morphology and insect behavior.

The concepts of symmetry handled in this manuscript are different with those that botanists are familiar with, and a bit more complex I have to acknowledge. Many references are made to the works of Yoland Savriama, and symmetry is, in this manuscript, conceived as a concept in crystallography. I would find useful for the readers to include, perhaps as an appendix, an additional figure explaining clearly the symmetric variation, the lateral and transverse asymmetry, etc.

Other comments and suggestions:
l. 55: replace “hermaphrodite […] population” with “hermaphrodite flowers are borne on different individuals”.
l. 108: add the information that the species belongs to family Celastraceae.
l. 118: write Euonymus in full.
l. 123: C4 notation: perhaps refer to the Schoenflies notation and add a citation. C4 usually means something totally different for botanists.
l. 141:  how distant were the 16 specimens from one another?
l. 210: “all four petals change their shapes”: I feel the passive voice would better fit here.
l. 279:  change “between” to “among”.
l. 295: insert “the” before “elongation”.
l. 369: the stamens are not strictly speaking “aborted”.
l. 429: change “intriguing” to “brand new”, for instance.

Are there any studies of floral development of Euonymus based on which you could compare the size of the meristem for each type of flower (female and hermaphrodite)? This could be another factor influencing the difference in corolla size between floral types.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: GYNODIOECY IN THE COMMON SPINDLE TREE (EUONYMUS EUROPAEUS L.) INVOLVES DIFFERENCES IN THE ASYMMETRY OF COROLLA SHAPES BETWEEN SEXUALLY DIFFERENTIATED FLOWERS
Review round: 1
Reviewer: 2

Output:
Basic reporting: The paper is well structured and well written. The cotext related to corolla dimorphism of gynodioecious species is well explained and refereed by the relevant citation. However, I find some ambiguity in the explanation of geometric method. For example, in line 100-102 the definition of asymmetric and symmetric components is a bit unclear. The same is true for the line 103-105 where the author addresses the possible association between corolla shape variation and stamen reduction. These parts should be rewritten in a more clear way.
Experimental design: The research question is well-defined and the approach to answering the question is clearly explained. Methods are discussed with sufficient details.
However, in line 161, the transformation method 2 is lacking.
Validity of the findings: The conclusions are linked to the original research questions and often supported by underlying data or relevant literature.
In line 377-379, the author mentions that “the smaller size and greater asymmetry of purely pistillate corollas might be related to the lower selection pressure on female flowers for visual attractiveness to insect pollinators.” For me, the opposite seems more plausible. The smaller size and greater asymmetry which is most likely a result of change in B genes function lead to lower attractiveness for insects.
Additional comments: The author conducted a comparative geometric study of corolla shape between hermaphrodite and female flowers of a gynodioecious species. The study is well-designed, structured and written. My main comments are mentioned in the other sections. I recommend the acceptance of paper after minor revisions.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: A CONVENIENT DIAGNOSTIC TOOL FOR DISCRIMINATING ADULT-ONSET GLUTAMIC ACID DECARBOXYLASE ANTIBODY-POSITIVE AUTOIMMUNE DIABETES FROM TYPE 2 DIABETES: A RETROSPECTIVE STUDY
Review round: 1
Reviewer: 1

Output:
Basic reporting: the aim of the present paper was to develop a convenient tool to discriminate adult-onset GADA-positive autoimmune diabetes from type 2 diabetes in patients with newly diagnosed diabetes. This retrospective cross-sectional study was conducted into n=152 GADA+, and n=358 T2DM.
The Authors identified five major factors discriminant functions: BMI, age at onset, TG, HDL-C, and HbA1c. BMI <23 kg/m2 was the most important factor, followed by TG <98 mg/dL, HDL-C ≥46 mg/dL, age at onset <30 years, and HbA1c ≥ 8.6%.
There are several points that should be clarified.

Major points.
1.	The introduction section could be more exhaustive including also the mention, in addition to  GADA, of  the other islet autoantibodies used to identify LADA patients (IA-2, ZnT8...) and also to the possibility that LADA patients are overweight and obese.( please refer and mention a recent review, i.e. Buzzetti R e at Nat End. Rev, 2017).

2.	The Authors reported that they have transformed all the continuous variables with statistical significance. They should specify the reasons why. Only the investigated variables not normally distributed should be transformed. Did they use a test to verify the normality of their data? They should report which kind of test they used.

3.	The Authors reported, as an important limitation of the study, that GADA were not measured in the type 2 diabetes patients, but in materials and methods they affirmed that patients with GADA titer ≥1.0 U/mL were excluded from type 2 diabetes. Could they clarify this matter? if GADA were not measured in type 2 diabetes this could be  a limitation of the study.

4.	Could the Authors specify more clearly the which were the exclusion criteria for the selection of the patients in their study? They reported that patients with an excessive alcohol consumption were excluded. How they evaluated the alcohol consumption?

5.	The Authors should show the ROC curve figures and not only the tables relative to the ROC curves..

6.	They reported that BMI <23 kg/m2 was the most important factor to discriminate GADA+ patients from type 2 diabetes. This cut off of BMI is very low. Considering such a low cut off there is the risk of not be able to identify correctly  patients with LADA. Many papers reported that LADA patients could often be overweight or obese. Recently Hjoirt et al found the importance of overweight/obesity and the risk of LADA (Diabetologia. 2018; 61(6): 1333–1343.

7.	Furthermore and more relevant, this low BMI and the young age of onset suggest a T1DM diagnosis! How the Authors can exclude that a substantial group of patients, particularly those in 20-29 age range could be classical T1DM? How did they differentiate LADA from T1DM? Which were the diagnostic parameters to identify a patient as having LADA instead of T1DM?

8.	Could the Authors specify using their diagnostic tool (BMI, age at onset, TG, HDL-C, and HbA1c) the number of patients with high GADA titer and low GADA titer identified.

9.	Some patients used statins and this treatment  could modify the lipid profile, therefore the two groups should be adjusted for use of statins.

10.	The Authors conclude that routine tests in diabetes care should be used to establish a convenient, low-cost tool that may assist in the early identification of adult-onset GAD+ autoimmune diabetes in clinical practice. However, to date, due to the broad heterogeneity of LADA, islet-cell antibodies measurement remains essential in order to decrease the number of misdiagnosis of diabetes. The Authors should underline this point in the discussion section.
11.	The Authors did not consider two relevant parameters that were took into account by Fourlanos et al in their tool: the presence of other autoimmune endocrinopathies in the LADA patient and a family history for autoimmune diseases. The Authors should mention why, also considering the high prevalence of autoimmune diseases in LADA patients, please refer to the following study Zampetti S et al JCEM, 2012 .

Minor points
1.	Line 154 replace “GAGA” with “GADA”.

2.	They should replace “sex” with “gender” in the text and table.
Experimental design: see above
Validity of the findings: see above
Additional comments: see above


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: A CONVENIENT DIAGNOSTIC TOOL FOR DISCRIMINATING ADULT-ONSET GLUTAMIC ACID DECARBOXYLASE ANTIBODY-POSITIVE AUTOIMMUNE DIABETES FROM TYPE 2 DIABETES: A RETROSPECTIVE STUDY
Review round: 1
Reviewer: 2

Output:
Basic reporting: The topic of the manuscript is to test the hypothesis that GADA-status can be precisely predicted by measuring standard clinical data such as BMI, age at onset of diabetes and standard biochemistry such as HbA1c and HDL.

It is acknowledged that GADA-measurement is not accessible in all settings. However, predicting GADA positive people is not a purpose of it-self. The clinical question to ask is whether the determined variables can predict insulin-dependency as good as (or even better than) than can GADA-measurement. The paper does not provide this answer nor discusses the topic.
Experimental design: A major issue is that the patients who did receive a GAD-measure are not necessary representative for the overall diabetes population unless all newly referred patients had the measure performed in the full study period. One may anticipate that GAD was taken primarily in the patients with higher suspicion of having and autoimmune diabetes.
Validity of the findings: The findings should be tested in an independent population to allow us to test the validity of the finding
Additional comments: The discussion section is much too long and not focused on the topic of the manuscript. For instances line 186-196 discussing mechanisms of insulin resistance is irrelevant
The age cut-of is somewhat arbitrary. GAD-positive patients in the present study will include young lean typical T1D patients whom are diagnosed in the absence of DKA.
L 143: “This study included a total of 510 patients, 152 in the GADA+ group and 358 in the T2DM 144 group”. This information is already given in the method section
145: GAGA+ group: correct typo
Overall I find the paper of modest interest. If the journal accepts short communication the results will be suitable for such a publication


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: A CONVENIENT DIAGNOSTIC TOOL FOR DISCRIMINATING ADULT-ONSET GLUTAMIC ACID DECARBOXYLASE ANTIBODY-POSITIVE AUTOIMMUNE DIABETES FROM TYPE 2 DIABETES: A RETROSPECTIVE STUDY
Review round: 2
Reviewer: 1

Output:
Basic reporting: the authors have answered all the questions raised
Experimental design: it is fine
Validity of the findings: it is fine
Additional comments: the authors have answered all the questions raised


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: A CONVENIENT DIAGNOSTIC TOOL FOR DISCRIMINATING ADULT-ONSET GLUTAMIC ACID DECARBOXYLASE ANTIBODY-POSITIVE AUTOIMMUNE DIABETES FROM TYPE 2 DIABETES: A RETROSPECTIVE STUDY
Review round: 2
Reviewer: 2

Output:
Basic reporting: The comments and concerns raised has been met satisfactorily
Experimental design: The comments and concerns raised has been met satisfactorily
Validity of the findings: I still believe that the manuscript would improve significantly if the suggested algorithm for predicting GAD-positivity was tested in an independent population. I know that is a lot of work, but as it stands the results are not readily implementable.
Additional comments: The comments and concerns raised has been met satisfactorily, although rather defensive.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: PHYLOGENOMIC ANALYSIS OF 589 METAGENOME-ASSEMBLED GENOMES ENCOMPASSING ALL MAJOR PROKARYOTIC LINEAGES FROM THE GUT OF HIGHER TERMITES
Review round: 1
Reviewer: 1

Output:
Basic reporting: The paper is well written and concise with good-shaped (supplementary) tables and figures. English text is excellent. I just list some minor suggestions and questions:

1. Please italicized phylum names (e.g., Fibrobacteres, Proteobacteria...) as the authors did for other taxonomic ranks (e.g., class Clostridia, family Lachnospiraceae...).

2. L233-234: It is difficult to follow why “genomes were recovered among the most abundant phyla from different gut compartments and diets” indicated "a good coverage of the diversity among gut compartments and host diets". Please explain more.

3. L236: "...which can be considered as abundant (Delmont et al. 2018)."
	In my opinion, what is "abundant" depends on the diversity of the sample. Anyway, I think that the authors can remove this description. One does not need to declare "0.23" is "abundant" or not.

4. L238: "..abundant members..."
	Here, "members" is ambiguous. Readers might regard it means "species". For example, "The present dataset includes major taxonomic groups of termite gut microbiota..." seems better.

5. L247-251: "...a significant and negative relationships between the number of metagenome-assembled reads and the MAG relative abundance..."
	Did authors mean here the relationships between the total number of metagenome-assembled reads in a sample and the mean MAG relative abundance of the sample?

6. L364-365: the class Chitinispirillia corresponds to TG3 subphylum 1 and Chitinivibrionia to TG3 sybphylum 2 (please see Sorodokin et al 2016 Frontiers Microbiol).
Experimental design: This paper describes the outline of comparative metagenomic analyses including binning up to 589 high to medium quality prokaryotic genomes (MAGs). This study with such abundant MAGs should be the basis for further detailed studies of termite gut microbiota, which is represented mostly by uncultured lineages. The experimental and analytical procedures follow standard methodologies and enough described.
Validity of the findings: The descriptions are based on their data adequately analyzed and previous findings. No  problems are found in "Results and Discussion".
Additional comments: I am looking forward to reading further analyses on these MAGs.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: PHYLOGENOMIC ANALYSIS OF 589 METAGENOME-ASSEMBLED GENOMES ENCOMPASSING ALL MAJOR PROKARYOTIC LINEAGES FROM THE GUT OF HIGHER TERMITES
Review round: 1
Reviewer: 2

Output:
Basic reporting: This manuscript is the first to do massive reconstruction of a large number (589) og MAGs from termite guts, spanning termites with different feeding habits. The authors significantly improved the initial automated assignment, frequently up to the genus level, recovering MAGs from most of the termite-specific clusters in the radiation of, e.g., Planctomycetes,
Fibrobacteres, Bacteroidetes, Euryarchaeota, Bathyarchaeota, Spirochaetes,
Saccharibacteria, and Firmicutes, which to date contained only few or no representative
genomes. The dataset is an important genomic resource and contributes substantially to populating the tree of life for gut-associated microbes. It furthermore will likely become an important backbone for a range of different follow-up studies that have been limited by the lack of genomic knowledge on termite bacterial gut members.

The structure, writing and figures in the paper are good, the referencing sufficient, and the language excellent.
Experimental design: The research questions are clear, the methods appropriate (and even extending beyond what many current studies with similar aims do) and the approaches and findings are important for research.
Validity of the findings: The presentation of the results and discussion of the paper is good and thorough, focusing on the more descriptive angles and less on the evolutionary histories that led to the observed patterns, nor their implications for termite-gut bacteria evolution. However, this makes the study solid, void of too much speculation, and very useful as a genomic resource paper, which is a major aim for the authors.
Additional comments: No additional comments