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: CHANGES IN PRECIPITATION MAY ALTER FOOD PREFERENCE IN AN ECOSYSTEM ENGINEER, THE BLACK LAND CRAB, GECARCINUS RURICOLA
Review round: 1
Reviewer: 1

Output:
Basic reporting: 1.	BASIC REPORTING
-	Clear, unambiguous, professional English language used throughout.
o	While the writing is general strong, the authors introduce numerous moments of ambiguity and breaks in flow that are both problematic and easily fixed. I recommend a substantive, line-by-line review of the current report with renewed attention to these weak points many (most?) of which I point out marginally in the ms.
o	I make multiple suggestions for potentially improved clarity and flow throughout the entire paper from abstract to figure legends. I was unable to comment on the pdf tables/figures themselves but attempt to make a few comments in the associated legends.
o	There are some key issues I have related to grammar and word choice. While these issues represent significant (and sometimes critical) flaws in presentation, nearly all are easily fixed.
	E.g., The use of the term “single preference” in regards to the single-item food tests is an oxymoron. No such thing. As such, the word preference can be replaced with “food consumption amount” or some such.
	E.g., The grammatical conjunction of “lettuce, apple and fish” (with only one comma) means something entirely different than “lettuce, apple, and fish” (with two commas). In the first example, the crabs have two options: lettuce OR apple and fish. In the second, they have three.
	Little details like these two emerge throughout the entire paper including the legends and make the reader work far more than we should to develop a tenuous understanding of what it is you want to convey.
	I flag most of these moments in the document and, thus, I hope that they are easily found and fixed.

o	STYLE-wise, the authors’ affinity for starting sentences with “This” or similar when “this” can refer to any one of several possible takers from the previous sentence introduces unnecessarily ambiguity.  Again, “this” is easily fixed.
o	Some of your sentences (marked) float in that they have neither connection to what precedes or succeeds them. Again, a few cleverly places transitional words and its fixed.
o	You may consider adding more verve to your writing by replacing the verb “to be” (e.g., is, are, were) with more powerful verbs (after all, any verb is more powerful than “to be” in all but the most intentional of circumstances).

-	Intro & background to show context. Literature well referenced & relevant.
o	The authors generally review the literature well and use a professional tone and style to illustrate several important elements that support the research to be reported. That said, I make many comments throughout about ways to further improve this intro.

-	Structure conforms to PeerJ standards, discipline norm, or improved for clarity.
o	The structure is conventional and, thus, form-fit to the industry’s standards. However, there is a missing subsection of the methods that I feel needs to be added: The study system description. I explain in the margins of the paper in far more detail than here just what I feel is missing. In summary, we need to know all relevant aspects of both the crabs (addressed in moderation elsewhere in the paper and thus easily moved to this new section) and the island forest ecosystem (including climate etc.) to understand better the experimental design and results/discussion to follow.  In summary, I find this to be an essential addition to the revision.

-	Figures are relevant, high quality, well labelled & described.
o	I would say that several of these figures are quite (perhaps, overly) complicated. I also believe that some of them are incorrectly presented. For example, I have been taught that when one finds significant interaction effects, then the individual elements of the interaction (although also statistically significant) are NOT presented as such.  Once the interaction is considered significant, the individual components can no longer be considered significant in the absence of the interacting co-variable. (If a significant interaction was NOT found, then (and, I have been taught, only then) would one present the factor in isolation. (table 4)
o	Additionally, I find the regression analysis to be unconvincing as we are looking at essentially flat line regressions with huge amounts of noise. My recollection about the use of p-values in regression analyses is that huge samples sizes (like they are presenting) can make even the most biologically INsignificant findings, statistically significant.  For this reason, some stats programs even refuse to calculate the p-values for such regressions (even tho it is easy to do) simply because they know that such p-values can throw off well-intended researchers into making a type 1 error.  I think this problem emerges within figure 7.
o	I feel that for nearly all figures and tables, additional context and description in the legends would be in order.
o	I also question the need for so many tables and figures as some of the information (e.g., table 1) could be easily incorporated into the text.  I recall that energetic content of the food was a detail specifically missing from the results text that I felt should be included (not sure I am recalling correctly but you can check).
o	I was hoping to see df for table 4 (or at least mention of sample size in the legend) for table 4 and similar. I was skeptical of the highly significant results for all factors and made me wonder about the potential likelihood of pseudo-replication (see more on this potentially serious concern below).

-	Raw data supplied (see PeerJ policy).
o	I cannot find the raw data (nor do I feel I need to see it).  I looked for it using the links supplied but they did not lead me to the raw data. I have no opinion about this one way or the other.
Experimental design: KEY: Prompts from the editors in bold.  My additions in bold italics.

2.	EXPERIMENTAL DESIGN
-	Original primary research within Scope of the journal.
o	To the best of my knowledge, this ms complies completely with the journal’s requirement for original primary research. I am assuming that the ms falls within your journal’s scope because I was asked to review it. Sorry, I am not yet familiar with your journal so cannot speak more without more effort But I have NO yellow flags of any sort on this point.

-	Research question well defined, relevant & meaningful. It is stated how the research fills an identified knowledge gap.
o	Largely yes. I find the authors’ work to be impressive in its intensity and scope but somewhat unpersuasive as to its relevance to natural circumstances either present or future. My main concern is about the powerfully contrived nature of the overall design which was highly manipulative and largely if not wholly artificial. (Although the authors do make a case for using novel food items not found in nature.)
o	The research imposes several hypotheticals some of which are more plausible (e.g., dehydration) or less plausible (e.g., unnatural food choices). Then, the researchers combine these distinct factors in ways that further compound the hypothetical aspects of the work.  Thus, I would have found this all more persuasive if naturally available foods were used instead.  I would have found it even more persuasive if field experiments were used to corroborate the lab manipulations.  However, that said, I feel that such work must begin somewhere and this early effort reflects a strong early go at the topic albeit from one extreme of the natural vs controlled spectrum.
o	Regarding the authors’ efforts to make a case that such research is relevant in the face of a drying and warming climate, I feel that they did a stand-up job. Well done because even a skeptic about the validity of lab manipulations—like me—found the argument somewhat compelling.

-	Rigorous investigation performed to a high technical & ethical standard.
o	Yes, in most ways but see below RE ethics.
o	They were thorough sometimes to the extreme in their work and design and I was regularly impressed with their attention to detail in design. They were rigorous in their data gathering (both in scope and detail) and analyses.
o	That said, I have a few lingering and important issues;
	Their presentation of their highly technical efforts left several key holes for their readership to fall through. As such, I frequently did not understand key elements of their methods and findings despite my dedicated attention during this review. Thus, I feel that it is the presentation of their work (rather than the work itself) most in need of revision.
•	For example, understanding the correct samples sizes of the crabs in the foraging experiments was, for me, nearly impenetrable. Thus, I could not fully comprehend (nor comprehensively review) these core elements.  That is just one of many such examples.
•	As such, a substantive, line-by-line revision (particularly of the methods section) will markedly improve the paper’s accessibility for an interested readership. All that said, the corrections are mostly quite simple. I suspect that they mostly reviewed the work amongst one another and, thus, were unable to reveal ambiguities that jump out of the page for a first-time reader. As one of these first-time readers, I was (I have to admit) rather reeling from the number of unanswered questions I had about process, purpose, design, etc. Again, all of this is easily (and essentially) repairable.
	Regarding ethics, they suggest that they are in compliance with some oversight effort but we cannot deny that the research amounted to a catastrophically difficult time for many of the crabs and, to be transparent, I admit that I struggled, personally, with this. So far as I can tell, 200 or more crabs (again, this part is unclear so I cannot be sure) were subjected to capture and captivation in unnatural conditions (e.g., wire cages). A subset of those individuals (again, unclear as to how many but over 150, I think) were subjected to various levels of dehydration. A subset of those were subject to levels of dehydration to the point that they became unresponsive (but revivable, I think).  And, 18 crabs (pretty sure) were placed in ice water to kill them so that they could be measured for dry weights and for use in what I felt was a somewhat non-essential efforts to calculate consumption amounts as a function of body weight.  I don’t feel that they well defended the need to gather dry body weight of the crabs as essential.
•	They might have killed one crab at a time to calculate wet wt – dry wt until the graph began to plateau at which point they would get the accurate value they sought, while, potentially, killing far fewer crabs in the process.
•	I think the collection of feces from the experimental subjects as they became increasing dehydrated could have revealed important physiological insights to their stress.  Testing the feces for moisture (wet wt – dry wt) might have been useful.
•	So, in confess, I was personally troubled by some of the methods used and not well defended because of their lethality. That said, they claim to have been in compliance and I do not doubt them.  (I just don’t find the laws of compliance all that worthwhile.)

-	Methods described with sufficient detail & information to replicate.
o	No. I feel that the report’s greatest weakness lies with the sheer number of points of critical ambiguity inadvertently built in (or left out) of the text. I have, perhaps, 50 or so questions about design details, data presentation details etc., which, in this current draft, remain unanswered or unclear (for me).
	The bad news is that I feel that without great clarification of these essential elements that pepper a majority of the ms, this paper should not be published.
	The good news, is that all of these concerns (well, most of them as some of them cut to the design itself) can be easily fixed through additional information, clarification of the information already provided, and, in some cases, proper use of grammar.
o	I suggest that they go line-by-line through the methods section and address all the questions that arise during/after a first-time reading of the materials.
o	That said, they had a huge amount of information provided and the authors should be commended for presenting a complex design and analysis in great detail.  However, given the complexity of their work, even more (far more) details are needed.

-	IMPORTANT ISSUE (NOT PROMPTED): Potential design flaws.
o	I am skeptical of the design of the food consumption trials (erroneously called “preference” trials) as well as the actual food preference trials (three types of food offered). In particular, I think we need to know how/when/if the food replenished during the trials. Was the food replenished regularly? Was the food allowed to desiccate over time? What sort of behavioral ecology was going on when males, females, juveniles and adults were mixed (i.e., did the juveniles or females eat less preferred foods due to the dominance of the larger adult males)?  In other words, I can imagine (but do not now know due to the incomplete description of the methods) potentially fatal flaws to this design that would render it unusable at worst or in need of major moderation of the interpretation of the results at best.
	For example, in the three-food trials, findings revealed that lettuce was not preferred compared to the apples and fish, and apples were eaten a lot until about day 4 of dehydration. Then apple eating dropped off.  Q: Did they eat fewer apples because there was no more apple left? Because their deteriorating condition led to a switch to fish from apple?  Because the apples became desiccated? These details that (so far as I recall) are missing from the methods description, radically alter the interpretations of the findings.
o	Pseudo-replication? When 14 crabs are placed into a bucket with a food source from which they eat or don’t, can we really reference this trial as having a sample size of 14?  I don’t think so lest we engage in pseudo-replication (of Stuart Hurlbert fame). Rather, because the crabs cannot be assumed to be acting independently of one another when confined in close circumstances and feeding from the same sources and, thus, they violate the assumptions of the statistical tests later performed.  As such, it appears to me that the sample size of that particular trial is 1 (for which there were 14 crabs involved). Therefore, this potential problem is far less easily fixed than the ones mentioned above and I think this should be further examined but this design (albeit not sufficiently well described in this current draft) feels like a classic case of pseudo-replication.  Perhaps the other reviewers caught this too?
Validity of the findings: 3.	VALIDITY OF THE FINDINGS
-	Impact and novelty not assessed.
o	(I don’t understand the negative angle of this prompt.) So, let me just say that the authors stated (inferred) well the potential impact and novelty (to a lesser degree) of their findings. If anything, they pushed the limits of this inference given the highly controlled and unnatural design they created. Thus, I feel this work is a good early step to assess the potential relationships between dehydration regimes and novel food consumption patterns.  So, if anything, I might tone down the inferences a bit, but I don’t think this is an issue that floats to the top for their next revision.
o	See comments in section immediately above about potential design problems and analysis problems.

-	Negative/inconclusive results accepted.
o	I don’t find persuasive their conclusions illustrated by figure 7 that presumably correlates food “preference” (poorly chosen term for a single-diet option treatment) and crab size (dry weight). I find these data to reveal absolutely NO relationship between the two variables yet the authors (citing the statistically significant results that I am claiming to be a possibly spurious artefact of the large sample sizes and their effect on the regression). I want to be clear that the authors seem to be more knowledgeable about statistics than I am so my confidence is lower than otherwise. But, I believe that they are making a Type 1 error here in which they are claiming a significant outcome where none exists (not biologically significant at any rate). If I am wrong, forgive this intrusion.
o	See my marginal comments for other issues.

-	Meaningful replication encouraged where rationale & benefit to literature is clearly
stated.
o	Again, this prompt confuses me a bit but their replication was, in general excellent so long as they did not engage in pseudo replication.  See my comments above regarding pseudoreplication.

-	Data [are] robust, statistically sound, & controlled
o	See above for concerns about pseudoreplication and lingering questions related to their food availability regimes (not described).  All of this is mentioned above in the section called “important issue”.

-	Speculation is welcome, but should be identified as such.
o	The authors are solid on this. They are generally modest in the phraseology of their extrapolations and assertions but bold in their desire to make them in the first place. I find this to be a strength of the paper as it makes for provocative reading.

-	Conclusions are well stated, linked to original research question & limited to supporting results.
o	Generally, yes. Except they bring up energetic content of the food which, so far as I could tell, was barely raised in the ms and sometimes equated with “nutrients” but without (as I recall) defense as nutrients and energetics (caloric content) are distinct so far as I know.  This should be checked and, if I am correct, easily fixed from abstract to legends.
Additional comments: Review of Changes in precipitation may alter food preference in an ecosystem engineer: the black land crab, Gecarcinus ruricola written by Iain J. McGaw,Travis E. Van Leeuwen, Rebekah H. Trehern, and Amanda E. Bates.

Reviewer: Peter Sherman, Prescott College, Arizona.

This study attempts to predict how individuals of a population of Caribbean black land crabs might respond to predicted future climatic changes with a focus on decreased rainfall and/or humidity. The authors hypothesize that food consumption rates and food type choice might be altered by increasing stress associated with advancing dehydration. To test their hypothesis, the authors create a serious of experiments that both dehydrate (or not) individual crabs while offering the crabs three different types of novel foods that differ in water, energetic, and nutrient contents. While the authors choice to provide the crabs with unnatural food items reduced markedly the potential to infer about presumed future ecological events, this study may provide an early step at addressing the larger issue. The authors claim that given the potential for land crabs to behave as ecosystem engineers, such alternations to their ecology, in general, and foraging behaviors, more specifically, might add additional impacts to forests anticipated to be experiencing their own ecological responses to pending drought stress.

The authors deserve recognition for that sheer amount of work that this document represents and their overall attention to detail in their design. I commend their creative use of triangulation (of methods) and their fearless and interdisciplinary approach to tackling the provocative problem. While I find the work to be impressive in both its scope and depth, I argue that the current draft of the manuscript to be sufficiently in need of additional work that I strongly recommend a massive revision prior to reconsideration. Given the number of issues that I raise marginally in the document as well as below, I suspect that their substantive revision warrants additional professional review to make sure that all needed (and many recommended) modifications are made. I would not accept the paper for publication unless the revisions are made and accompanied by a highly detailed, comprehensive point-by-point (for the major points) description of how each point was addressed (either in compliance or disagreement).  Importantly, many (but not all) of my critiques can be addressed rather easily. All of them, I think can be addressed—none of them are, to the best of my knowledge, fatal.

Please know that despite the critical and rather comprehensive nature of my review, I acknowledge that I might have missed several things (e.g., some of the statistics I did not understand) and many of my comments might have implied some missing information that the authors had provided (but I somehow missed or forgot etc.)  All that said, however, I did give this my full attention for many hours and, thus, I think it would be wise for you to consider that if I am genuinely confused about something, that perhaps you (as the authors) might find a way to clarify the work even more than it already is. As the reader, I am prepared to do my fair share of the work to understand your work. But, I feel, that ultimately and generally the onus lies with the authors to go to whatever extend necessary to maximize the clarity of their prose. And so, I provide you this critique with the greatest of respect and empathy for how it feels to have to go back and revise the document one more time. Good luck with this. I look forward to seeing it published.

-	General comments
o	On the one hand, this is an impressive paper crafted by impressively trained authors who have provided a highly sophisticated and detailed account of a provocative research topic. The sheer breadth and depth of their efforts are impressive and should be commended. The detailed statistical analyses (some of which extend beyond my own training) appeared to be robust and well developed.  The work, although highly contrived, provides an early understanding of how land-based, marine invertebrates might adapt (or not) to a drying climate. All in all, an impressive effort worthy, eventually, of publication.
o	However, many substantive omissions of key details hamper my ability to fully understand (and, thus, review) the work. While most of these are fixable with another substantially altered draft (or more) following my many marginal prompts for additional clarity and accuracy, I worry about some design elements (mentioned above) that might be more problematic.
o	Regarding potential pseudoreplication, even if the n = 14 crabs trial is reduced to a n = 1 scenario due to the inter-dependence of the crabs foraging together in a single container over time, I think that they can fix this by reanalyzing the data and revising the findings accordingly.
o	Even if the data emerge to be statistically insignificant under that dire reduction in sample size, the biologically meaningful information remains the same (e.g., the crabs don’t like the lettuce as much as the apples etc.). So, they lose much of their statistical prowess but they maintain the main message that, when foraging in a forced group setting, less lettuce is eaten than similarly forced groups of crabs eating, say, apples.
o	For the real preference trials of three food choices in one tank, then the sample size issue become even more confounding as the individual crab’s foraging choices are absolutely influenced by one another and the very notion of independence in this scenario is ecologically untenable especially if the forage was not replenished every day (or more) or provided ad libitum.
o	And so, in summary, the paper requires a major revision of which a vast majority of the many need easily fixed. And, for those more elemental issues raised just above, the merits of the paper to be published depends, in large part, on their responses to these critiques.  However, even under the worst case scenario where my concerns are well-founded, the authors might still be able to redeem the situation by simply modifying their analyses (redoing or eliminating many of them) but allowing the underlying (more important) biology to remain up front and center as these potential concerns don’t so strongly impact the biological findings as the statistical ones.


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: CHANGES IN PRECIPITATION MAY ALTER FOOD PREFERENCE IN AN ECOSYSTEM ENGINEER, THE BLACK LAND CRAB, GECARCINUS RURICOLA
Review round: 1
Reviewer: 2

Output:
Basic reporting: Line 153: Give species names
Line 249: This is results not methods.
Line 348-351: I had to read this a few times to understand what it was saying. Clarify.
Line 354: SD? SE?  Clarify throughout, e.g. in Fig 3
Experimental design: The only time repeated measure is OK is if time is not expected to have an effect (which it clearly is here as the crabs will get more dehydrated over time).  When that’s not expected to have an effect, the analyst can include time as a factor to check that there is no significant interaction between the factor of interest (in this case food type) and the repeated measures.  But here the authors are specifically interested in time so they couldn’t do that.  So they needed to either (a) include crab as a random factor (which would require a lot more measurement of each individual at each time period and doesn’t really solve the problem anyway) or (b) have different individuals for each food x time treatment (which would require a lot more crabs).   I think the authors have done (a; Line 289-292), but I don’t really follow how “each crab potentially feeding on the 3 different food items” creates replication, so need more details before I could give a blessing.  If it doesn’t work, then their results are uninterpretable. :(  In which case the only thing they could do with the data they have is to only look at the start and end results, subtracting 8 d from 0 d for each individual and then having a single data point per crab that then gets used in analyses just looking at food type.   But I’m getting ahead of myself, since I don’t follow what they’ve done….
Line 158: Why sex them if they were then randomly allocated?  If sex not mentioned elsewhere, just leave out.
Line 183: And because researchers are asleep?  Also, I understand that respiration experiments are difficult to do when the critters are all active, which is why night doesn’t work, but you don’t actually say that, which would be confusing for readers not familiar with these sorts of experiments. So make that link, if that is what you mean by avoiding the night time experiments.
Line 212-217: Those crabs: http://2.bp.blogspot.com/-7hAXgapnsg0/U9mYRThh2ZI/AAAAAAAACnU/KQATCUiR3eQ/s1600/andy.jpg.  Also, do you mean goad, rather than agitate?
Line 235: I think this a long bow to draw, there’s more to the food items that them being new to the critters and the water content.
Validity of the findings: Line 170: So we don't know that's it's just water loss?  It's just change in mass, right?  I think the text should be written throughout to reflect this nuance.  I don't think it really affects the conclusions, but better to be rigorous in saying what you mean throughout.
Line 318-340:  This is just a big slab of qualitative and anecdotal information backed up by a single photo.  Not sure what to do with this as I can’t just accept what’s being said.  So this slice of natural history might as well not even be in the paper. :/
Additional comments: Line 160: Except the eight that you dehydrated until they became moribund (thanks for giving me the chance to learn and use a new word!) at 14-16 days. Or is this just a funny way of saying that there aren't any Canadian and Bahamian care protocols for crustaceans?


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: CHANGES IN PRECIPITATION MAY ALTER FOOD PREFERENCE IN AN ECOSYSTEM ENGINEER, THE BLACK LAND CRAB, GECARCINUS RURICOLA
Review round: 2
Reviewer: 1

Output:
Basic reporting: x
Experimental design: x
Validity of the findings: x
Additional comments: x


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: CIRCULAR RNAS AS POTENTIAL BIOMARKERS AND THERAPEUTICS FOR CARDIOVASCULAR DISEASE
Review round: 1
Reviewer: 1

Output:
Basic reporting: English renders proofreading
Experimental design: N/A
Validity of the findings: N/A
Additional comments: Review on the article titled “Circular RNAs as potential biomarkers and therapeutic for cardiovascular disease” by Wang et al., 2019
The authors present a comprehensive review encompassing implication of circRNA in cardiovascular disease  (CVD) factors. The review presents the mechanistic  insights of circRNA biogenesis, genetic aspects of CVD factors and a review of current research on implication of circRNA on CVD.
While the structure and style of review is acceptable, the numerous grammar problems renders English proofreading. Also, biogenesis of circRNA description is currently non-transparent and should be reworked according to the notes below.
Notes:

1.	P. 6 (43-44) “In human genome, noncoding RNAs (ncRNAs) account for the majority of transcripts and only 1~2% are protein-coding genes” – please reference this statement. Protein coding genes comprise approximately 40% of the genome. The transcripts usually correspond to full-length genes. If this is even true in terms of numbers/percentage, it should be referenced correctly, not comparing transcripts to genes (“…transcribed from protein coding genes”).
2.	P6( 44) housekeeper ncRNA – housekeeping RNA? Never heard of these, please reference/explain.
3.	First paragraph of the intro should present circRNA as well, while currently doesn’t.
4.	The approximate numbers/percentage of each RNA class should be presented in human genome within the abovementioned paragraph.
5.	P6 (63) “since long”?  <=>“for long time?”
6.	P8(98): liner = linear
7.	The first paragraph of circRNA biogenesis chapter (98-109) should extensively reference Fig. 1 with corresponding indices (a-d) for each instance, in contrast of referencing the figure at the end of the paragraph. This paragraph is currently not transparent and is hard to apprehend. It should be extended for more clearly manifested/illustrated  details and with the intense referencing to the figure, since it’s a vital chapter for further reading. It can be done by using Fig. 1 as a directory to consequently describe each variant, extensively explaining (not deciphered in Fig. 1. legend) terms like ecircRNA,  EIciRNA.
8.	P9 (112-113): “can stop (prevent?) circRNA formation  by melting the stem structure” Relating to above: where is stem structure in the figure? Is it fig 1 b? It should be referenced to the figure in this instance.
9.	P11(186): “is exist”
10.	Fig 1 should preferentially be arranged for a vertically oriented page, with 4 cases presented separately per string(s). The typeset should be larger. All abbreviations in legends should be presented therein. Fig. 2 has some of denotations placed on the dark red/violet background that are not readable, typeset should be also made bigger, at least in legend.


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: CIRCULAR RNAS AS POTENTIAL BIOMARKERS AND THERAPEUTICS FOR CARDIOVASCULAR DISEASE
Review round: 1
Reviewer: 2

Output:
Basic reporting: The article discusses recent reports on the assessment of the biological significance of circular non-code RNAs and their possible diagnostic and prognostic value as markers of cardiovascular diseases. This topic is very relevant. In recent years, numerous experimental and review articles have been published, discussing the role of circular non-coding RNA in the regulation of transcription and translation processes in cells. Several review articles have been published, discussing the possibility of using circular non-coding RNAs as markers.
The article is written in good English. Article written in professional English.
The review is of broad and interdisciplinary interest that meets the requirements of the journal.
Experimental design: The research methodology corresponds to a comprehensive, unbiased coverage of the subject.
Literature is cited adequately.
The text of the review is well structured and organized logically in the form of coherent subsections.
Validity of the findings: The text is well thought out and consistent with the objectives for the purposes set out in the introduction.
In addition to the Conclusion, the article presents the Limitations section, where the factors limiting the use of non-coding circular RNA as biomarkers for diagnosis and diseases outcome prediction.
Additional comments: Сomments on the text of the article are given in the attached file.


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: CIRCULAR RNAS AS POTENTIAL BIOMARKERS AND THERAPEUTICS FOR CARDIOVASCULAR DISEASE
Review round: 2
Reviewer: 1

Output:
Basic reporting: The authors addressed issues of circular RNA research, which definitely is a novel and important research area. It is relevant not only to cardiovascular disease area, but to cancer research, and many others.
Experimental design: Acceptible
Validity of the findings: N/A
Additional comments: As of now I have no major concerns to the 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: CIRCULAR RNAS AS POTENTIAL BIOMARKERS AND THERAPEUTICS FOR CARDIOVASCULAR DISEASE
Review round: 2
Reviewer: 2

Output:
Basic reporting: The review is of broad and interdisciplinary interest that meets the requirements of the journal.
Experimental design: The research methodology corresponds to a comprehensive, unbiased coverage of the subject.
Literature is cited adequately.
The text of the review is well structured and organized logically in the form of coherent subsections.
Validity of the findings: The text is well thought out and consistent with the objectives for the purposes set out in the introduction.
Additional comments: A few more comments on the text of the article are given in the attached file.


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: MODULATION OF STRESS RESPONSE AND PRODUCTIVE PERFORMANCE OF LITOPENAEUS VANNAMEI THROUGH DIET
Review round: 1
Reviewer: 1

Output:
Basic reporting: This Manuscript needs to be improved before publication: some suggestions are listed below

There is no information in introduction about type of diet (composition) which has already evaluated on shrimp cultured in or submitted to low salinity condition. This would be useful to introduce the choice of experimental diet composition used in this study.
There is also no mention of probiotic interest in diet to face stressful conditions.
The use of “global means” all along the “Result” part does not facilitate the comprehension. The readers have to calculate all cited values (as these mean values are not include in the corresponding tables) to understand what the authors mean.
Experimental design: Some important details on the experimental conditions are missing in the text (the supplementary data were useful to find these missing details in manuscript):

#Line 82-92: Initial shrimp weight at the beginning of differentiate feeding period; The number of replicate per treatment (salinity and diet); the number of shrimp used (total and per treatment)

#Line 101-106 and Table 1: More details on diets composition and ingredients are required : origin and proportion of carbohydrates, origin and concentration of probiotic and yeast (commercial or lab strain, genre and species if known), different mix composition). The diet formulae are not complete, sum of ingredients cited below 100%. What was the percentage of free amino acids in diets?

#Line 128-131: How long after stress, are the shrimp harvested and sampled?
Validity of the findings: The authors showed that an adapted diet improve survival, protein conversion rate and productive performance when cultured at 6psu. The shrimp capacity to face stressful condition is also improved by this adapted nutrition: the shrimp energy status is improved after 42 days of culture whatever the salinity

Parts of "Discussion" have to be reviewed :
#Line 325-352: This part of the discussion on the CBH and its impact on productivity is a bit long, while there is no information on CBH level in the experimental diet of this study. This part should be reduced if additional data on the diet formula is not added in Material and Methods (cf previous comment).
#Line 352-354 : Even if « Probiotic » is  a small part of the experimental diet, it would be interesting to specify in the Mat & Max, what type of bacteria and yeasts were added and for which purpose. Especially if these microorganisms could have contributed to assimilate CBH.

Data about survival to additionals acutes stresses (escape and hypoxia) could complete interestingly the results obtained in this study.
Additional comments: Pay attention:  "psu" in not written correctly in different places; “37 psu” is sometimes replaced by “35 psu”
Line 167: “growth of” can be deleted


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: MODULATION OF STRESS RESPONSE AND PRODUCTIVE PERFORMANCE OF LITOPENAEUS VANNAMEI THROUGH DIET
Review round: 1
Reviewer: 2

Output:
Basic reporting: The present manuscript is well structured, the objective is clear, English is well written, the bibliography is adequate, I suggest that some of the tables could be transformed into a figure to facilitate the visualization of the results.
Experimental design: The present research is original because it integrates the effect of environmental stressors with diet and in response it measures several biochemical and aquacultural parameters. The research question is well defined. The methodology used is described in sufficient detail and can be replicated. Also in the part and results it is suggested to group them as they are too long and it is tedious to follow them.
Validity of the findings: The  literature cited is considerad that adequate and covers the subject treated. The impact of this research is related to shrimp management from the aquuacultural point of view by the focus of the evaluated parameters. In the part of the statistical analysis I suggest the authors use the arcsine transformation in the parameters that they describe and that are in percentages because to apply the ANOVA in that type of data they must be transformed by that function since otherwise they would be violated the assumptions with which the statistical test works. The conclusions are considered adequate in relation to the results obtained
Additional comments: In the part of the statistical analysis I suggest the authors use the arcsine transformation in the parameters that they describe and that are in percentages because to apply the ANOVA in that type of data they must be transformed by that function since otherwise they would be violated the assumptions with which the statistical test works.
Also in the part of results it is suggested to group them because as they are presented they are too long and sometimes repetitive.
On line 330 it is suggested to change the prayer to. In a recent study Wang et al (2015) used isoenergetics diets with different protein: CBH ratios of 26-30%: 30-14% were compared with the highest growth at 3 psu was observed with a diet consisting of 19% CBH and 34% protein.


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: COMPARISON OF MICROBIAL COMMUNITY STRUCTURES IN SOILS WITH WOODY ORGANIC AMENDMENTS AND SOILS WITH TRADITIONAL LOCAL ORGANIC AMENDMENTS IN NINGXIA OF NORTHERN CHINA
Review round: 1
Reviewer: 1

Output:
Basic reporting: The English used in the ms is acceptable for its understanding although some corrections are recommended
Introduction should include more information on the state-of-the-art of microbial communities and soil properties. This information can also be used to help in the discussion of the ms.
The ms is difficult to read. Perhaps, most of the Results section could be removed because it is summarized in the Discussion. The Discussion, after selecting the results in it, is quite limited and should be improved. Similarly, the provided Conclusions are mostly a repeated text of Results and should be more elaborated.
This is a descriptive ms with provide data on microbial communities and soil properties at two time-periods (7 and 15 months) for several soil treatments using different sources for the amendments.
Experimental design: The topic of the ms fits within the Aims and Scope of the Journal.
The objective of the ms is the comparison of different soil amendments on microbial communities and soil properties. Nevertheless, it is not well defined how the provided results fills current knowledge gaps. The ms present some data which are potentially of interest but they are poorly analyzed and no meaningful conclusions are clear.
Methods are described but some improvements can be appropriate. Most characteristics of soil and amendments could be provided in a Table so that the text can be reduced and simplified. It is not clear the quantity of soil and the amendments mixed in the treatments; for instance, does the weight of soil corresponds to dry weight?.
I have a major methodological concern to this ms. The addition of a N and P source to soil and amendments. I understand that supplementing soil with different amendments is aimed to maintain or improve soil nutrients and properties so that plant growth is enhanced and soil impoverishment is avoided. However, the authors add a N (urea) and P (KH2PO4) source to their treatments. The addition of extra N and P sources besides the amendments could hide the effect of different amendments on soil properties and above all on microbial communities because potential restrictions on these nutrients might be masked.
The authors mention "pyrosequencing" in lines 103 and 205. However, they do not use this technique, rather they use Illumina sequencing (line 226). Please, correct because the Illumina platform does not perform pyrosequencing.
Please, avoid reporting Bray-Curtis distance and mention Bray-Curtis dissimilarity which is the correct way. This is an approach to beta-diversity which is not mentioned in the ms.
Please, avoid reporting Bray-Curtis distance and mention Bray-Curtis dissimilarity which is the correct way. This is an approach to beta-diversity which is not mentioned in the ms.
Please, correct OTU throughout the ms instead of OUT.
Statistical procedures do not look to appropriated due to the inconsistencies reported by the authors repeatedly throughout the ms.
Validity of the findings: The final outcome of the performed comparison is far from being clear. The data are there but the authors should analyze them more in depth. Perhaps, different, more appropriated statistical treatments are needed, perhaps some methodological factors contribute to mask (see above) clear effects of the amendments. It would be desirable to provide some meaningful conclusions to the author’s set of data beyond a simple (difficult to read) description.
In order to improve the readability of the ms, I would recommend to simplify the text and report in writing the important observations which will contribute to important discussions and conclusions. For example, most of the Results (correct Results and not Rusults) could be deleted. Most of the important stuff is included (repeated) in Discussion. Removing the Results, and marking Results and Discussion the current Discussion, the ms will be much more readable. The Discussion is also awaiting for improvements. Based on a more in depth Introduction, the Discussion could built up and add new knowledge to the state-of-the-art providing information beyond a simple description of results. A more informative Conclusions section could be written. The statistical analysis of the results does not look to appropriated judging for the “inconsistencies” that the author report repeatedly as pointed by the authors (line 365). Perhaps, the authors, after a detailed analysis, could provide with explanation for those data. An advice would be to simplify the results; for instance, the use of one diversity index would make the analysis easier than using four different indices.
Care must be taken when approaching potential physiological features to 16S rRNA sequencing data. Cyanobacteria should be considered, at first, as primary producer (photosynthetic) microorganism common in soils rather than as a copiotroph (line 447). A priori, one should expect that the availability of inorganic N and P sources might rule their development.
A major result of this set of data could be that pH, SOC and TN could be the most important parameters driving microbial communities during the amendments of soil (lines 399-400).
Tables 4, 5 and 6. What is according to the authors the difference among Relationships and  Correlations?
Figure 1 shows differential distribution of treatments but Figure 2 shows no major differences among the different phyla for each treatment. The authors should consider if these differences are significant.
Additional comments: The authors should try to analyze in detail their results in accordance with current state-of-the-art so that informative conclusions, adding up to current knowledge, can be provided. Different potential suggestions have been provided 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: COMPARISON OF MICROBIAL COMMUNITY STRUCTURES IN SOILS WITH WOODY ORGANIC AMENDMENTS AND SOILS WITH TRADITIONAL LOCAL ORGANIC AMENDMENTS IN NINGXIA OF NORTHERN CHINA
Review round: 1
Reviewer: 2

Output:
Basic reporting: This manuscript is an original research paper investigating the impact of different soil amendments on the diversity and community structure of soil microbes (Bacteria and fungi), and the soil chemical properties that shape microbial diversity and community structure. Fundamentally the study is solid and well design.  The authors should revise the reference list carefully, cross-check with the text as some reference is cited in the text and missing in the reference list. The referencing style should be checked carefully in-text as well as in the reference list and corrected according to the journal instruction to authors. The spelling for some author’s names in the reference list is not correct. In general, the manuscript is well written. However, I have raised some minor issues that the authors should address.
Experimental design: Experimental design is solid and well done. The multivariate statistic methods and statistical tests used are suitable for the study. I’ve got no criticism in this regard.
Validity of the findings: Besides evaluating the influence of amendments on the diversity and community structure of the soil microbial diversity at two point times (after seven weeks, and after fifteen weeks), the authors compare the influence of two local organic amendments (cow manure, corn straw) versus a woody organic amendment (poplar branches) on soil microbial community structure. Very little works have been done previously on that aspect in the field of microbial ecology. The study is, therefore, important as it is a welcome contribution that advances the field of study, and contributes to the body of knowledge. In general, the manuscript is well written. However, I have raised some minor issues that the authors should address.
Additional comments: The authors should revise the reference list carefully, cross-check with the text as some reference is cited in the text and missing in the reference list. The referencing style should be checked carefully in-text as well as in the reference list and corrected according to the journal instruction to authors. The spelling for some author’s names in the reference list is not correct. In general, the manuscript is well written. However, I have raised some minor issues that the authors should address.

Introduction

Line 75: Please, check the year of publication for the citation. It is listed as (Fernández-Gálvez et al., 2009) in the text, but as “2012” in the reference list (line: 529).

Line 85: Please, check the year of publication for the citation. It is listed as (Dai and Chu, 2012) in the text, but as “2010” in the reference list (line: 519). Also, replace “and” with “&” in the citation of interest.

Line 87: The citation (Jia et al., 2013) must be in italic

Lines 99-100: The citation (Barrios, 2007) is not listed in the reference list

Material & methods

Line 147: add authority to “Populus alba”.
Line 148: add authority to “Zea mays”.

Line 196: Please, check the year of publication for the citation. It is listed as (Nelson et al., 1982) in the text, but as “1996” in the reference list (line: 577).

Line 201: Replace “and” with “&” in the citation “(Tran and Simard, 1993)”

Line 213: The citation (Weisburg et al., 1991) must be in italic

Line 215: The reference must be cited as “(Gardes & Bruns, 1993)”

Line 239: “(Edgar, 2013)” must be in italic

Line 242: OTU and not “OUT.”

Line 283: Please, clarify the meaning of “CM was also separated from CM”

Lines 284 and 284: Please, indicate clearly if it is either (Fig. 1a) or (Fig. 1b), or both?

Line 316: I suggest that the output result of db-RDA be represented by a “biplots” as well, showing the soil chemical properties that significantly influence the bacterial and fungal communities, respectively. The plots of interest can be made in “vegan” after the analysis.

Discussion

Lines 357 and 358: Add a “comma” after “ et al.” to the citations (Francioli et al.,2016; Sharma et al., 2017)

Line 366: Replace “and” with “&” in the citation “(Hartmann and Widmer, 2006)”

Line 367: OTU and not “OUT”

Line 368: The citation “Calleja-Cervantes et al. (2015)” is not listed in the reference list

Lines 404 and 405: Add a “comma” after “ et al.” to the citations (Lauber et al., 2009; Rutigliano et al., 2014)

Line 423: Add a “comma” after “ et al.” to the citations (Zeng et al., 2016; Lladó et al., 2017; Zeng et al., 2017)

Line 428: The year of publication for the citation “(Tian et al., 2014)” is “2014” in the text, but “2015” in the reference list (line: 599)

Line 441: The same citation is written as “(Tian et al. 2015)”. Please, check!

Line 442: Add a “comma” after “ et al.” in “(Yashiro et al., 2016)”

Lines 452 and 471: Replace “and” with “&” in the citation “(Větrovský and Baldrian, 2013)”

Line 472: The citation (Berg, 2014) is not listed in the reference list

Line 513: REFERENCES and not “EFERENCES.”

Reference List

Line 514: “Caporaso et al.” was not cited in the text and should be deleted in the reference list

Line 519: Replace "and" with a "comma" in “Dai G and Chu W. 2010”

Line 520: The journal’s name “Forest Resources Management” must be in italic

Line 521: Add a full stop after “2013” in “Edgar RC. 2013”

Line 529: “2009” or “2012”? Please, check the year of publication carefully

Line 533: Add a full stop after "RB" in “Fierer N, Bradford MA, Jackson RB 2007.”

Line 577: “1982” or “1996”? Please, check the year of publication carefully

Line 608: Remove the brackets around “2006.”

Line 613: The Author's name “Voříšková” is misspelt either in the text (line 452) or in the reference list. Please check!


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: COMPARISON OF MICROBIAL COMMUNITY STRUCTURES IN SOILS WITH WOODY ORGANIC AMENDMENTS AND SOILS WITH TRADITIONAL LOCAL ORGANIC AMENDMENTS IN NINGXIA OF NORTHERN CHINA
Review round: 2
Reviewer: 1

Output:
Basic reporting: The ms has improved considerably and it is almost ready for publication
Experimental design: Experimental design has been reexplained and it is satisfactory
Validity of the findings: A great improvement is observed in the last version of the ms
Additional comments: Just two points for the authors to improve in their ms:
- lines 258-259: Please, correct: ... 10,510 sequences were considered per bacterial community and ...
- Conclusion. lines 437-458. The conclusions sections can be significantly reduced. for example:
Bacterial and fungal community compositions changed significantly for different amendments. Non-traditional organic amendments such as woodchips (treatment PB) exhibited similar effects to corn straw treatments (CS). Both PB and CS treatments influenced soil communities differently that the most traditional amendment using cow manure (CM). Soil organic carbon, total nitrogen and pH were the most important factors driving microbial communty structure. Woodchips should also be considered as a viable soil amendment for future applications.


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: LEAF FUNCTIONAL TRAIT VARIATION IN A HUMID TEMPERATE FOREST, AND RELATIONSHIPS WITH JUVENILE TREE LIGHT REQUIREMENTS
Review round: 1
Reviewer: 1

Output:
Basic reporting: No comment
Experimental design: No comment
Validity of the findings: No comment
Additional comments: This manuscript describes a study of naturally occurring juveniles of 19 woody plant species in a temperate rain forest and the relationships between foliar traits (structural and nitrogen concentration) and leaf lifespan in shade and in canopy gaps.  Leaf lifespan among species was more strongly correlated with LMA in canopy gaps, while leaf lifespan among species was more strongly correlated with the species’ minimum light requirement in the shade.  This is a well written manuscript with clear goals, and well-analysed appropriate data.  The study shows the need to understand plasticity in foliar traits.  It also shows that the range of leaf lifespan and foliar traits represents a narrower range than in the humid tropics.
Since there are neither data on soil nutrients, no evidence is presented in this manuscript to support a view that seasonality (or lack of it) overrides soil fertility (line 262) as a driver of short leaf lifespan.  The data in Lusk et al. (2015, per line 265) is at the whole site scale and may not be applicable for species that are local in their distribution (e.g., lines 127, 226).  I think the manuscript could drop lines 260–267, and instead state that the interactions between local soil nutrient availability and light with respect to leaf lifespan and nutrient use efficiency remain to be determined.
Minor points:
Line 55: It seemed an oversight not to include references from New Zealand or Australia.  If this is strictly about juvenile leaves, I am unaware of published data, but if interpreted more broadly (cf. line 38; mostly about adult leaves), then consider citing Richardson et al. (2010) and references therein for New Zealand.
Line 56: Remarks about “northern continents” (line 55) are broadly true, but comparisons on leaf lifespans, LMA, etc. can also be drawn with broadleaved evergreen temperate forests of eastern Asia (e.g., Grubb et al. 1975, Kusomoto 1978, Nitta & Ohsawa 1997, Huang et al. 2007) (and subtropical broadleaved forests such as those of Macaronesia).  Consider also (cf. line 58) the largely evergreen rain forests comprised of conifers in the Pacific Northwest of North America.
Line 65: Rozendaal et al. (2006) is also very relevant here.
Line 104: Plagianthus regius is the species (for which P. betulinus, not betuloides, is a synonym).
Line 104: McGlone et al. (2004, cited) lists Fuchsia excorticata as fully deciduous, but notes two contrary references about its deciduousness in the northern part of its range; it would be as well to note why this study regards the species as semi-deciduous.  McGlone et al. also consider Aristotelia serrata (line 134) as semi-deciduous, which seems to fit the text at line 133 and the data about leaf lifespan in Table 2 (0.7 year length in the shade, 0.6 in the open, also line 224).
Lines 138, 143: What of herbivory by dama wallaby and red deer? (cf. https://www.doc.govt.nz/globalassets/documents/conservation/threats-and-impacts/animal-pests/bay-of-plenty/okataina-wallaby-report.pdf, which lists several of the species in Table 1 as consumed by wallaby or deer).
Line 266: Replace “somewhat” with the mean (± SE) percentage longer in the shade.
Typos:
Line 241: Hyphenate “novae-zelandiae” (as in Table 1)
References
Grubb PJ, Grubb EAA, Miyata I 1975 Leaf structure and function in evergreen trees and shrubs of Japanese warm temperate rain forest I. The structure of the lamina. Botanical Magazine, Tokyo 88, 197–211.
Huang J, Wang X, Yan E 2007 Leaf nutrient concentration, nutrient resorption and litter decomposition in an evergreen broad-leaved forest in eastern China. Forest Ecology and Management 239, 150–158.
Kusomoto T 1978 Photosynthesis and respiration in leaves of main component species. In: Kira T, Ono Y, Hosokawa T (eds) Biological Production in a Warm-Temperate Evergreen Oak Forest of Japan (JIBP Synthesis, Volume 18), pp. 88–98.  Tokyo University Press, Tokyo.
Nitta I, Ohsawa M 1997 Leaf dynamics and shoot phenology of eleven warm-temperate evergreen broad-leaved trees near their northern limit in central Japan.  Plant Ecology 130, 71–88.
Richardson SJ, Peltzer DA, Allen RB, McGlone MS 2010 Declining soil fertility does not increase leaf lifespan within species: evidence from the Franz Josef chronosequence, New Zealand. New Zeaalnd Journal of Ecology 34, 306–310.
Rozendaal DMA, Hurtado VH, Poorter L 2006 Plasticity in leaf traits of 38 tropical tree species in response to light; relationships with light demand and adult stature. Functional Ecology 20, 207–216.


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: LEAF FUNCTIONAL TRAIT VARIATION IN A HUMID TEMPERATE FOREST, AND RELATIONSHIPS WITH JUVENILE TREE LIGHT REQUIREMENTS
Review round: 1
Reviewer: 2

Output:
Basic reporting: I have provided a list of suggestions that I hope are found useful to the authors.
Experimental design: I have provided a list of suggestions that I hope are found useful to the authors.
Validity of the findings: I have provided a list of suggestions that I hope are found useful to the authors.
Additional comments: The manuscript Leaf functional trait variation in a humid temperate forest, and relationships with juvenile tree light requirements is an interesting study on the relevance of considering light environments when working on the role of functional traits on plant strategies. However, there are several aspects of the manuscript that were not very clear to me, I hope that my suggestions help and improve this study.

L 49. Can you include a reference to support this statement?
L 50 This statement is confusing, “less well-documented” than?  Are the tropics very well-documented? Could you provide a reference to support this statement?
L52-53 This study is a simulation that considers traits of only one (very common) pioneer species from Australia… could you include more references, as this is a rather strong statement and the whole idea does not seem to be supported with literature (I am referring to the previous statements that did not include references).
L53-55 Is this a comparison among evergreens only? If you exclude any deciduous do you still get such a variation?
L61 Hallik et al. 2009 is not on evergreen forests only, this work includes deciduous species and actually reports a correlation between leaf lifespan and shade tolerance.. I am missing the point of this last sentence (L58-61).
L62.
L67 change “than” for that
L67 Lusk et al. 2008b main conclusion is related to thickness and herbivory and this study was made in tropical Australia, how can it support the argument of LMA running counter to inherent variation in shade tolerance?

L 86 “infilled” is written in different font
L109 a space is missing between “(LMA) is”

L112 Indeed thickness does not correlate but can you add a reference that supports that density does? Kitajima´s paper reported the importance of thickness but not density, and again, they (Kitajima and Poorter) highlighted the role of herbivory.
L115 It would be more correct to say “some tropical forests”.. both studies were carried out in the tropics and Poorter 2009 actually includes dry tropics.

L116 (In Table 2 says “Life lifespan” instead of “Leaf”)

L129. Please provide more information on leaf lifespan calculations. How can you calculate leaf lifespan of leaves that live 3 or 5 years by following only 12 months? If I understand you followed “fully expanded leaves” on a branch, but how do you know how old are these leaves? You can tell how many leaves die, but how do you determine the mean and the error of these calculations?

L 142 why depending on leaf size? Why is this relevant?
L 181-187 I understand the value of considering phylogeny, but this is never mentioned in the introduction, so why to do this test? Could you include something in the introduction that could then provide a possible effect of phylogeny on your questions and results? What could we expect if there is a significant phylogenetic signal? How would differences in shade- or sun- traits be?

L194- 205 these statements require statistical support, how do we know that something is “more marked”, that ranges are “narrower”, that a trait responded “more strongly”? The way Table 2 is sorted makes it even harder to figure out the point the author is trying to make here, why not to run a “t test” between sun- and shade- leaves? In that way all this sections would be properly supported.
L199-201 Is this a species-level or community-level comparison?  How do you calculate the 59%?

L 209 The usage of bold and bold/underlined I confusing, as the table already has a lot of information, I recommend to change to *0.05 (barely significant) and ** for 0.01 (significant)

L 211-212 But MLR is negatively correlated -0.74 and -0.84 (in bold and underlined) with LL in sun- and shade- leaves respectively, how can you state that this was “especially for shaded leaves? Given this I really do not see support for the statement “leaf lifespan was thus positively (shouldn´t it be negatively) correlated with species’ shade tolerance”

L 213 Light requirements were correlated with LMA in -0.47 (P<0.05) in sun-leaves, but in -0.66 (P<0.01) in shade-leaves, hence, it seems fair to me to say that LMA is actually positively correlated with shade tolerance.

L219-220 These statements seem a bit misleading, as one species is deciduous with very short leaf lifespan, and the 7-year species… without those extremes, variation is actually 4 fold in shaded environments and 3.5 fold in light environments. The 7.7 LL has a standard deviation of 5.4 so do you think that is valid to consider it?

L 220-222 I am sorry but I don´t see the connection between this sentence and the previous one, where is the connection between the tropics and your results.

L 219-234 I am sorry again, but I am bit lost with the point, are you trying to discuss why are there not many deciduous or short-lived leaved species neither in your assemblage or in Chile?  Is it not very clear to me and there are studies that have already discuss this, one is Matt´s study, already cited in your paragraph.

L235-245 As previously mentioned, in some aspects of the introduction, in the tropics, thickness has been correlated with herbivory (Kitajima & Poorter 2010) or the study was made in a tropical/diverse region where herbivory may play a relevant role (Lusk et al. 2010). I think that the point you are making here is lacking the relevance, or irrelevance, of herbivory in temperate forests?

L247-255  Very interesting indeed, however, I think that information on soil nutrient availability is lacking, can nutrient availability affect LMA, leaf lifespan or dry matter content? I can see that there is some discussion in the following paragraph, however,  soil nutrient availability previous studies also in New Zealand have shown the importance of soil nutrient availability for shade tolerance and plant growth strategies.  See Coomes et al. 2009. J. Ecology (/doi.org/10.1111/j.1365-2745.2009.01507.x)

L276-279 I think that this is a very interesting point, but I don´t this is supported with data, temperate humid forests in Australia and Chile have fast-growing tall pioneers, how do you account for that? Or is this an irrelevant point?

L279-282 I may have misunderstood your results, but in order to be sure that I did not, I would like to see t-tests for each species comparing LMA values (gap vs. shade) as I wonder, given the standard deviation values, if indeed there is support for your conclusion.


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: LEAF FUNCTIONAL TRAIT VARIATION IN A HUMID TEMPERATE FOREST, AND RELATIONSHIPS WITH JUVENILE TREE LIGHT REQUIREMENTS
Review round: 2
Reviewer: 1

Output:
Basic reporting: This manuscript is of a high standard four all four criteria for basic reporting.
Experimental design: This manuscript meets all four criteria of experimental design.
Validity of the findings: The manuscript meets all standards set to give confidence in the validity of the findings.
Additional comments: This revised manuscript describes a study of naturally occurring juveniles of 19 woody plant species in a temperate rain forest and the relationships between foliar traits (structural and nitrogen concentration) and leaf lifespan in shade and in canopy gaps.  It is a clear, well-written study, generally improved by revision.

In my earlier review, I commented that without data about soil nutrients, I felt that the manuscript did not have sufficient evidence to say that the scarcity of deciduousness in the New Zealand woody flora “is more a reflection of weak seasonality than soil fertility” (line 265 of the current manuscript).  In response, I think the author makes a reasonable case that the material should stay.  As the author notes in response comments, there is an issue of scale-dependence in assessing whether soil fertility drives leaf lifespan, and I would prefer that this was noted in the manuscript.  I would prefer that an additional caveat sentence was inserted in the paragraph beginning at line 264 to say that interactions between local (i.e., within-site, such as Okataina) soil nutrient availability and light with respect to leaf lifespan in the New Zealand woody flora remain to be determined.

Another comment is that the expression of full deciduousness in widespread New Zealand trees is necessarily related to temperature, as shown for Aristotelia serrata (Dungan 2001) (cf. line 104).

Minor corrections
Line 91: insert “the” before “Department”
Line 99: No italics on “Cunoniaceae”
Line 307: Italics on Aristotelia chilensis
Line 393: a 2010 publication cannot be “in press”

Reference
Dungan RJ 2001 Whole‐tree winter leaf‐loss in wineberry (Aristotelia serrata, Elaeocarpaceae) is not related to mean air temperature. New Zealand Journal of Botany 39, 547–550.


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: EFFECT OF ISOLATION ON COAT COLOUR POLYMORPHISM OF POLYNESIAN RATS IN ISLAND SOUTHEAST ASIA AND THE PACIFIC
Review round: 1
Reviewer: 1

Output:
Basic reporting: I’m not a native speaker myself, but I had the impression that the text might profit from being proof read by a native speaker, as I think the use of words and grammar is incorrect in several places. Further, many sentences are incomplete and confusing.

The article includes sufficient introduction and background to demonstrate how the work fits into the broader field of knowledge. Relevant prior literature is appropriately referenced.

The structure of the article is conform to an acceptable format of ‘standard sections’.

The figures are relevant to the content of the article and of sufficient resolution. However, the Supplementary figures have no captions.

The submission is ‘self-contained,’ represents an appropriate ‘unit of publication’, and includes all results relevant to the hypothesis.
Experimental design: The research question is well defined, relevant & meaningful. It is not directly stated how research fills an identified knowledge gap, but this becomes evident from the introduction.

Rigorous investigation is performed to a high technical & ethical standard.

Methods are described with sufficient detail in most parts (others are specified in the general comments below) & information to replicate.
Validity of the findings: The data on which the conclusions are based is provided, as far as possible (no photographs from every single specimen are provided, but this is probably not feasible). The data is robust and statistically sound (as far as possible to tell, but I think more details should be provided, see below).

Conclusion are well stated, linked to original research question & limited to supporting results. However, I have several concerns regarding underlying concepts and limitations of the data, which are not sufficiently discussed (see general comments below).
Additional comments: The author investigated coat colour in a great number of Polynesian rat specimens from mainland areas and islands. The goal of the study was to investigate, if aberrant coat colour (e.g., white patches, lighter or darker coloration) occurs more often in the island populations than in the mainland ones, as a result of release from predator pressure and in analogy to domesticated animals. It was found, that aberrant coat colour occurs only on the islands, but hat this occurrence is not causally linked to island size, persistence time on the island, and number of predators and competitors. Instead, it is hypothesised that the observed changes are results of the proximity to humans and unintentional selection for tameness, limited gene flow, and genetic drift.

The research question is very interesting and worthwhile, as it uses the analogy of domestication and island evolution to elucidate fundamental connections between phenotypic traits and their mechanistic underpinnings. The sample is great, the data sampling seems sound, and the analyses appropriate. However, I think that shortcomings of the material should be discussed, as they are crucial for the validation of the results and the conclusions. In particular, the observed pattern of no aberrant colours in mainland specimens could be the result of a sampling bias (particular collectors prefer specimens with white patches over the “normal” ones) and/or a smaller sample size in the mainland group compared to the number of specimens from the islands would automatically increases the probability to find aberrant coat colours in island specimens. Such discussion of limitations are lacking.
Further, I feel that there is a bit of a confusion among concepts, particularly “commensalism”, “commensal pathway of domestication”, and “island evolution”. I am sure that the author knows the differences, but it is confused in the text. For example, commensal house mice (Mus musculus) on the continent are not considered to be selected for tameness along the commensal pathway of domestication. Instead, M. musculus is considered to be a commensal wild species (see e.g., the study on white patches in wild mice selected for tameness by Geiger et al. 2018). On the other hand, the author describes the Polynesian rats on the islands as human commensals that are therefore selected for tameness along the commensal pathway of domestication. This inconsistency between the concept of “commensalism” and “domestication along the commensal pathway” between these species should be discussed.  (It might help to clarify things if you would further describe the nature of the commensal relationship between the Polynesian rats and humans. Are they living in a closer relationship than e.g., house mice and humans? Are there more/closer interactions? I am sure this is difficult, if not impossible to answer. But such issues should be mentioned and discussed to be able to fully evaluate the results). Second, I think that domestication and island evolution, although potentially resulting in similar phenotypic traits, are fundamentally different processes. On islands, there is normally no selection for tameness, in contrast to domestication. According to the neural crest cell hypothesis of the domestication syndrome (Wilkins et al. 2016), changes of neural crest cell migration would therefore not play a role in island evolution. Instead, and as also suggested by the author, non-adaptive factors (e.g., genetic drift) and predator release may play a role in generating the peculiar traits of mammals on islands. These differences should be more clearly described in the text. In the manuscript, it is stated at several places that the underlying mechanisms between the domestication and the island syndrome are the same (e.g., L. 18 – 19, 96 - 99).

Lastly and as stated above, many sentences are incomplete and confusing and should be edited.
Further, I’m not a native speaker myself, but I had the impression that the text might profit from being proof read by a native speaker, as I think the use of words and grammar is incorrect in several places.

These are my main concerns about the manuscript. Please find more detailed comments below:

L. 17 and throughout the text: “domestication syndrome” is sometimes accompanied by apostrophes and sometimes not. Please be consistent.
L. 18: “Simultaneous” implies a temporal component, which is not the case in this context. Maybe exchange with “concurring” or “co-occurring”?
L. 21: “their native range” instead of “its native range”
L. 22: It is not clear what you mean by “this”. Do you mean “aberrant coat colour has been found in …”? Specify please.
L. 24: I think that the convention is to write out numbers below 10. So this would mean “seven” instead of “7”. Further, do you mean “in the course of seven centuries since colonialization of these islands”? Please specify. Moreover, I think that this sentence is a bit misleading, as it implies that you were investigating rats from different time periods, dating back several centuries, which was not the case (?). Consider re-phrasing please.
L. 28 – 29: What is meant by “relaxed selection under commensality”? Less predation?
L. 49: include “and” before the last point of the list.
L. 52: in some cases there is evidence for an increased reproductive output. See e.g., Raia et al. 2003 (Evolutionary Ecology 17: 293–312)
L. 56: “melanisme” is without an “e” at the end
L. 61: Include a comma before “or”
L. 62 onwards: It might be interesting to mention e.g., the Falkland island fox as an island species that has previously been described as exhibiting domestication features (Clutton-Brock J, 1995, Science: 7-20)
L. 64: Replace square bracket in the citation.
L. 64: As far as I know, the term “domestication syndrome” has been used for the first time in animals by Wilkins et al. 2014 (Genetics, Vol. 197, 795–808) on the basis of a term which has originally been coined for crops. The cited works by Zeder do not use the term “domestication syndrome”.
L. 68: Why “amongst others”? As far as I know the only selection criterion was tameness/aggressiveness.
L. 77 – 78: This sentence is a bit weird and does not make much sense. It is particular changes of the neural crest cell migration, which might contribute to the characteristics of the domestication syndrome.
L. 83: The selected trait in this case is tameness. It might make it easier for readers to say so, instead of stating “traits that were directly or indirectly selected for”
L. 85: There is an “of” missing after “irrespective”.
L. 89 and 100: Please be consistent with brackets in the cases of the citations of the people that described the species.
L. 95: What do you mean by “spontaneous” evolutionary change in this context?
L. 96: Consider rephrasing the sentence to: “evolutionary changes in murids, which are typical for the domestication syndrome”.
L. 96 – 97: As already mentioned above, I don’t agree that the domestication syndrome and the island syndrome have a “partial developmental similarity”. The phenotypic results of domestication and island evolution are similar, as pointed out earlier during the introduction, but I don’t see why the developmental underpinnings should be the same. In the commensal pathway of domestication, there is selection for tameness and maybe alterations of the neural crest cell development. On islands, there is no selection for tameness and therefore probably not such a similar developmental link. I would therefore rephrase this sentence and not use the term “development”.
L. 101 – 104: This sentence is a bit confusing. Why should domestic rats develop depigmentation in parallel with other domesticates? After all they are domesticates already and you described earlier that depigmentation occurs in rats. I think this sentence is not needed at all.
L. 106 onwards: Is this species presently found on all these islands? Please specify the time period.
L. 109: do you mean it lives as “human” commensal? Please specify. Further, why do you list “scrubs” as an anthropogenic habitat? Could you please specify this further?
L. 111: What do you mean by “predator”? Are they regularly hunted by humans for food or as pests? Are they hunted with traps or otherwise?
L. 115 – 116: This sentence seems not complete: Compared to what? Further, you do not describe here why island area, which is analysed later on, is important in the context of the research question. Please add a background here.
L. 120-122: You refer to “specimen numbers” twice and to different Tables, Data S1 and Appendix S1? Which is the right one? Also, there is no file called “Appendix S1” or “Appendix S2” etc. Could you please provide unequivocal file names?
L. 124 and following: Would it be possible to indicate these island groups in Fig. 1 and its caption? It’s quite hard to follow you here.
L. 129 – 132: Could you provide references here, please?
L. 146: How did non-specialists evaluate this? Did you let non-specialists look over a certain portion of the furs? How do you define “significant” difference (usually, this term is only used for statistics)
L. 152: This white belly only occurring in the Flores population is very interesting in itself, in the context of the research question, isn’t it? It might be worthwhile to discuss this further.
L. 160 – 170: Could you please describe the statistical tests in more detail? E.g., how did you define the groupings for the tests and why did you use a Kruskal-Wallis Test for islands and a Mann-Whittney U test for island area?
L. 163: “evenly” instead of “evently”
L. 165 and otherwise: I’m not sure if I’ve seen this notation of a superscript “10” in log10 before. Isn’t it usually subscript or on the same level?
L. 173 -174: Do you mean Figure 2 instead of 3 here?
L. 174 – 176: I think I understand why you exclude the albino from the analysis (because albinism is not considered a part of the domestication and island syndromes). But please describe this further here or in the discussion, as it is quite important. Further, and as described below, I do not see the differences between the albino and the leucistic individual on the figures. Could you please specify?
L. 187: This sentence is incomplete.
L. 188: I do not understand this sentence. Do you mean that the difference of distribution of aberrant coat colour over the regions is not significant?
L. 201 and following discussion: There were many more specimens from islands than from the mainland. Could it be that there are more aberrant colour patterns on islands by chance? Please discuss why or why not.
L. 203 – 207: This sentence is somehow inconsistent: Do you argue that there is no correlation between aberrant coat colour and time in isolation because on the first and last colonized islands there was a similar amount of specimens with aberrant coat colour?
L. 208: Do you mean “predator” instead of “predature”? Where are these results?
L. 220 – 221: Could you please elaborate on why you think that this is an artefact? This is not clear from the text.
L. 225 – 226: It is not clear what you mean here. What do you mean by “insularity itself”? The influence of isolation on coat coloration via genetic drift and isolation? You might repeat here that all island rats are human commensals.
L. 239: Do you mean “particular” instead of “preferred” (which implies an intention)
Further: As far as I see, Geiger et al 2018 did not describe the nature of the colour polymorphism, which they observed in the mice. I mean, there is no quantification or description of the location of the white fur. They merely stated that there are white dots and patches of fur in some specimens. So it is not really clear if these patches resemble the ones in brown rats or the Polynesian rats and this comparison is therefore not feasible.
L. 244: For house mice, commensalism alone has not changed coat colour, but selection for tameness, i.e., a much closer relationship to humans than just the general proximity has probably resulted in coat colour changes (see Geiger et al. 2018). Do you think that this might be different in the Polynesian rats?
L. 245: The commensal pathway of what? Domestication?
L. 248 – 249: But you found that number of predators is not correlated with the occurrence of the aberrant coat colour. This sentence contradicts your results. Are there more predators on the mainland than on the island(s) with the highest number of predators? If yes, please present these numbers and discuss.
L. 255: Why is this (what exactly do you mean by “this”) not the case on Batam island? Because there are no predators anyway that would make such a coloration adaptive? Or because the vegetation/geology is different?
L. 262: What do you mean by “a higher degree of domestication”? I think these Polynesian rats are not domesticated, but commensal wild populations. Do you mean a greater number of characteristics that occur in the domestication and island syndromes?
L. 263: shift “observed” after “The” and before “colour polymorphism”.
L. 264: There is two times “distribution” in the list.
L. 303 – 304: Switch the positions of “further” and “might”
L. 313: delete “here”
L. 326: delete either “as” or “compared to”
L. 326 – 328: But you found a correlation between the colour polymorphism and island area and number of competitors. So this is contradicting. Do you mean a causal correlation?

Fig.1:
- Many geographic areas mentioned in the legend are not indicated on the map (e.g. Greater and lesser Sunda islands). It would help the reader a lot if you could indicate those on the map.
- “…Sulawesi, Moluccas etc.”: What do you mean by “etc.”? Specify please, or mention just the most important islands.
- Lines 4 and 5 in the caption: “after which” and “subsequently” mean the same and one of these expressions should be removed from the sentence.
- Consider splitting the second sentence of the caption in several sentences to make it easier to follow.

Fig. 2:
- What do you mean by “larger” geographic region? Consider just deleting “larger”
- Delete “Data for” in the caption title. It is not needed.
- There is something missing in the last bracket: “details of ventral (A) and dorsal skins (B-)”
- it is very hard to interpret the figure if it is in greyscale (printed).
- Would it be possible to provide a picture of normal coat colour here? This would make the comparisons more straightforward.

Fig. 3:
I do not see the differences between leucism and albinism that you describe in the materials and methods section. On what basis did you classify these specimens as albinistic/leucistic?

Citations:
The format is somewhat inconsistent. Publications with three authors are sometimes cited with all names in the text (i.e., x, y & z 2018) and sometimes abbreviating all names but the first (i.e., x et al. 2018). Please make this consistent according to the journal requirements.

Supplementary Figures:
- I did not find figure captions and could not follow the descriptions in the text. Could you please provide captions and/or indicate the patterns which the reader is supposed to see with arrows?
Further, it is quite hard to keep an overview over the many figures in the main text and the supplement. I would suggest to put also the supplementary figures in the main text and combine figures to make it easier for the reader to keep an overview. Raw data has been made available.


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: EFFECT OF ISOLATION ON COAT COLOUR POLYMORPHISM OF POLYNESIAN RATS IN ISLAND SOUTHEAST ASIA AND THE PACIFIC
Review round: 1
Reviewer: 2

Output:
Basic reporting: This interesting manuscript examines the distribution of aberrant fur color patterns in a widespread rodent across continental and insular settings. An impressive number of specimens was examined, with all relevant records of aberrant color patterns being from islands. I particularly like the link between the island syndrome and the domestication syndrome that the author makes. I have several specific comments, which the author could consider. See general comments for the author, below.
Experimental design: No experiment was conducted. The study simply compares continental and insular specimens of rodents, which is a valid and interesting scientific comparison in this manuscript.
Validity of the findings: I recommend analyzing the data with a different statistical method. See general comments for the author.
Additional comments: The manuscript is rather long, given the results. Some of the Introduction and Discussion could be pared down, particularly the rather lengthy discussion of the genetic basis of aberrant fur colors.

There is a potential collection bias in the specimens examined. I recall speaking with an old-style museum collector (long since deceased) of mammals, who said that they captured so many rodents at times that they would simply discard those that they considered to be of no interest. It is possible that such collectors fanning out across Southeast Asia and the Pacific islands would consider either wild type specimens or mutant specimens to be of no interest, thereby introducing a collection bias into the data base. I am not suggesting that it actually happened, but the possibility exists, and the audience perhaps should be aware of that.

The extremely small number of aberrant specimens would seem to render any statistical test just about unnecessary because of the low likelihood of finding any statistical significance. However, I suggest that the author could try a more powerful type of test, such as a linear or loglinear analysis of categorical data. Those tests also calculate a chi-square value, but they are based on maximum likelihood estimation and are typically considerably more powerful than a simple chi-square test, particularly if there are any cell sizes <5. There are two results where P<0.1, and a more powerful test may reveal significance, even with very small cell sizes.

On Line 99, the author states that a specific hypothesis was tested. It was tested only in a statistical sense because no experiment was conducted. I would use “address” or “examine” or some similar word rather than “test”.

There are a number of places where the grammar, spelling, and word usage could be improved. I will not edit the entire manuscript but will point out a few such places.

Line 53: “suit” should be “suite”.

Line 56: “melanisme” should be “melanism”.

Line 177: it is usually better to use “by contrast” rather than “on the other hand”.
Line 187 ends in mid-sentence, at least in my version of the manuscript.

Line 208: “predature” should be “predator”.

Line 228: something appears to be missing in that sentence.

Line 239: I would not use “preferred” in this context. “Specific” seems to be more appropriate.

Line 246: it is stated that the Polynesian rat is a human commensal where it has been introduced. I have captured many Rattus exulans on remote Pacific islands that are sparsely populated by humans, far from any human habitations or influences. While they may be frequent commensals of humans, they may often occur also in a completely sylvatic state.

Lines 297 and 299: I believe “mouse deer” should be “deer mouse”.

Line 322: “the other islands of our data set were not sampled.” I see only one author listed, so “our” should be “my”. The author should take full credit for assembling such an impressive data set.