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: CLIMATIC NICHE EVOLUTION IN THE VIVIPAROUS SCELOPORUS TORQUATUS GROUP (SQUAMATA: PHRYNOSOMATIDAE)
Review round: 1
Reviewer: 3

Output:
Basic reporting: Grammar and english used in this ms made it difficult to read, and needs substantial work and refining. Some paragraphs are normal length, others are overly long. Past and present tense is confused throughout. Specific scientific listing of species (i.e., using the full genus name at first mention and at the beginning of sentences, followed by abbreviation thereafter) is also not followed throughout. Most background is provided, but there are substantial sections of the introduction that are not necessarily related to the study. References seem appropriate. Some analyses are inconclusive and should probably be excluded. Otherwise structure seems good. I do not necessarily consider the conclusions drawn here to fit with the results.
Experimental design: The data here are largely from published sources, but ideas are original and interesting. The questions asked here are relevant and meaningful and if undertaken correctly would fill a research gap. In my opinion, the data are overly speculated upon and do not provide as concrete evidence for the conclusions drawn as is written - and nee to be toned down substantially. There are often details missing from the methodology that preclude a complete understanding of what was done, and would certainly preclude replication of this study by others.
Validity of the findings: I think the results here are overstated in the conclusions drawn - and there is substantial speculation regarding the conclusiveness of these results. Sample sizes are relatively low for power in the main tests used herein to draw conclusions, others of interest (i.e., DTT results) are not examined at all despite interesting patterns emerging from the data.
Additional comments: Specific comments: I am providing substantial comments here in order to assist the author in refining the ms to be suitable for publication.

Abstract:
Ln 1-2: Correct to "...explanation for the evolution of viviparity in reptiles. This hypothesis maintains...."
Ln 7: Correct to "...to test whether their diversification is ...."
Ln 8-10: Correct to "...We evaluated phylogenetic signal and trait evolution, along with reconstructions of ancestral climate tolerances, and did not find evidence in support of PNC in the ecological niche of the species group."
Ln 11-12: Correct to "...; we only found evidence of PNC in ...."
Ln 13: Correct to ..."Analyses of relative trait disparity through time..."
Ln 13-16: First it is stated that there is evidence for divergence associated with orogenic cycles, and no PNC - but then it is stated that this is not due to niche differentiation. These statements somewhat contradict each other - and I would argue that there is evidence herein that there is niche differentiation (i.e., the absence of conservatism)

Background:
Lns 2-43: this should be split into at least 3 paragraphs - not as a single paragraph
Ln 2: What does "actual" distribution mean? Are you referring to the realized or potential niche of the species?
Ln 8: Correct to "...are important for the speciation...". Also note: it is now commonly believed that divergent natural selection alone cannot lead to speciation without cessation of gene flow through allopatry to assortative mating. See (Cotto & Servedio 2017 AmNat 190 (5), 680-693; Servedio & Boughman 2017 Annual Review of Ecology, Evolution, and Systematics 48, 85-109 as a start)
Ln 14: Correct to "...that limit adaptation to climatic..."
Ln 18: Phylogenetic signal in climatic traits?
Ln 19: Correct to  "...random from a phylogenetic tree..."
Ln 21: Correct to "...have highlighted theoretical problems with the PNC concept and practical..."
Ln 24-25: Correct to "...questions are developed..."
Lns24-32: These arguments are confusing. I urge a rewrite for clarification of arguments.
Ln 33: The author suggests a topic is "barely known" but cites a reference from 1985. Have there been no recent developments in this field or at least a more recent paper citing the lack of this knowledge?
Ln34: Correct to "...interaction of these constraints..."
Ln 35: Correct to "..In this regard, viviparity..."
Ln 39-40: Which life history traits specifically?
Ln 44 Correct to "Viviparity among squamata..."
Ln 45 add a comma after the Blackburn references
Ln 46 Correct to "...viviparity in vertebrates..."
Ln 47: Correct to "...evolutionary studies regarding the relationship between niche evolution..."
Ln 48: Correct to "...Sceloporus. Sceloporus species are widely distributed in North America, contain..."
Ln 50: delete "for which there is"
Ln 51: Correct to "...species is available along with a comprehensive occurrence..."
Ln 53-54: Correct to "Given the hypothesis that the development of viviparity in reptiles is linked with the occupation of cool climates, this could lead to constraint in niche evolution among viviparous ..."
Ln 55: delete "organisms" after "model"
Ln 56: Correct to "...assess if niche evolution is phylogenetically constrained among viviparous.."
Ln 57-58: Correct to "...tolerances among species relative to their phylogenetic relationships fits with the PNC hypothesis; and 3) test whether the most.."
Ln 59: Are these the most important variables determining the extent of distributions? Please be more specific here.
Lns60-81: This paragraph should be either put in the methods, in a more concise format, for justifying species selection - or simply removed as most of the information presented here seems irrelevant to the paper.
Lns82-85: Correct to "...used phyloclimatic analyses that use occurrence data and bioclimatic variables to: 1) evaluate phylogenetic signal of bioclimatic variables comprising species niches,..."
Ln 86: Please define what variables are most important for?
Ln 88: Correct to "...calculate ecological niche disparity..."

Materials & Methods:
Ln 96: Correct to "...and mistakes in coordinates (i.e., points on the sea). To minimize..."
Ln 99: Correct to "...arc seconds). For environmental layers, we used bioclim layers at 30 arc seconds..."
Ln 100: Correct to "...1km) including monthly..." Delete s from "temperatures"
Ln 101: Correct to "...precipitation variables from the WorldClim.....(http://wwwwroldclim.org). We also used annual..."
Ln 102: Correct available to "variable" and put parentheses around the website.
Ln 104: Correct to "...clipped to limits of the distribution of all species in the group combined." This would allow for overprojection of species distributions, expecially for those with much narrower distributions relative to other species, impacting the validity of the results. A more appropriate method is to clip the layers to each species distribution and undertake MaxEnt analyses on each of these data separately. This will avoid overprojection problem.
Ln 116: Correct to "...relationships..."
Ln 119: Were those layers based on those that were important for all species or most important across species? How was this determined?
Ln140: Correct "achieved" to "performed"
Lns 143-155: This is the first time that it is mentioned that both the torquatus and poinsettii groups are being used. They are also listed as "former" groups in some places later on. This is neither clarified or explained and I feel should be mentioned before now in the text.
Ln 143: Correct to "...tree of the Sceloporus..."
Ln 144: Correct to "...group that resolved some..."
Ln 145: Delete being and add a comma after "studies"
Ln 146: make a new sentence after the references and change "they to Leache et al. (2016)
Ln 147: add a comma after the first "species"
Ln 148: Change "like" to "as the"
Ln 149-150: Correct to "...(2016), correpsinds to S. cyanogenys according to Martinez-Mendez de la Cruz (2007), therefore should not have affinity with S. serrifer..."
Lns151-155: I would be hesitant to take this statement as conclusive evidence of a misID given the stated paper is based on mtDNA alone and using different specimens (i.e., UWBM 6636 is listed in Leache et al (2016) using high-throughput sequencing while UTA-R-24004 is used for the Martinez-Mendez de la Cruz (2007) paper based on a few mtDNA genes). Are there morphological reasons to believe that the species' can easily be misIDed? What are the distributions of the two species'? Do they both occur in this region? These are relevant pieces of information that are not clarified here and make this statement as currently written inconclusive.
Ln 156: Correct to "...S. torquatus group"
Ln 157: Correct to "...we used sequences from four..."
Ln 160: add "the" before "grammicus", delete the "the" and change to "outgroup"
Ln 163: "outgroup"
Ln 163-164: Leache et al. (2016) actually finds 100% support for reciprocal monophyly of these groups - not "problems" as stated here. Therefore the provided reference here contradicts the authors statement.
Lns165-173: PartitionFinder would likely be a more appropriate choice of program to assess models of molecular evolution and data partitions here given these are largely concatenated analyses.
Ln 173: Correct to "...Phylogenetic relationships of members of the torquatus..."
Ln 179-180: Correct to "...inclusive parameter-rich model for analyses."
Ln 185: What does "edition" mean in this context?
Ln 193: Correct to "...the toquatus and poinsettii species groups..."
Ln 194: Correct to "...S. serrifer..."
Ln 199: Correct to "...Despite criticisms of the methods used to estimate PNC following..."
Ln 203: Correct to "...(BM) mode of trait evolution,..."
Lns 205-209: this is confusing as written and should be refined.
Lns199-242: This should be broken into at least 2-3 paragraphs
Ln 212-213:  Correct to "...with 1000 simulations, Pagel's.......using a maximum likelihood method."
Ln 223: Correct to "...to test for fit of the data to four alternative models of trait evolution: 1)..."
Ln 224: delete "as we pointed out models" and "the"
Ln 225: add "trait" before "evolution, and delete "of a trait"
Lns 223-232: Disentangling a listing of the four models tests and their definitions would assist in making this text less confusing.
Ln232: Correct to "...of trait evolution was assessed using log likelihood values and..."
Ln 233: Correct to "...AICc was deemed the better..... Additionally, to clarify our ability to distinguish..."
Lns 231-239: It is not actually clear which of these you used to determine the best model. Also given your sample sizes it is not surprising that you in fact have little ability to distinguish among models for any of these variables. Bio15 is the only variable for which there is a clear model that can be distinguished and it fits a BM model of evolution. See comments below about assuming the results of these analyses are conclusive.
Ln 243-245: What data are you using here?
Ln 247: adaptive shifts?
Ln 253: Correct to "...method uses a..."
Ln 255: Correct to "...also be indicative of speciation.."
Ln 256: move parenthetical statement to after Warren's I
Ln 257: Correct to "...), both of which range from......"
Ln 258-259: Correct to "...that are likely incorrect, where there were significant differences between values of the I statistic relative to Schoener's D...". Also - what are these unrealistic assumptions specifically?
Ln 260 Correct to "...for further analyses,..." Also - if there are problems with one, why use both, not just the more accurate one?
Ln 267-268 Correct to "...relates suitability scores within species distributions (from MaxEnt analyses) to..."
Ln 269: Correct to "...of each species for..."
Ln 273: How were weighted means calculated?
Ln 282: Do you mean pairwise distances in weighted average PNO's? How was this calculated? If distances you are basically analyzing the evolution of similarity and divergence in climatic variables. This is ok - but should be clarified here and detailed in the explanation of the results relative to the hypothesis being tested.
Ln 284: Correct to "DTT is calculated..."
Ln 286: add "trait" before "evolution"
Ln 287: Correct to "...subclade was plotted..."
Ln 289 Correct to "...indicate trait disparity is distributed..."
Ln 290: Positive values for MDI only infer early trait evolution if they appear early in the evolutionary history of the group, consistent with an early burst model. Therefore, these two aspects - divergence between subclades, and early evolution - are being confused here.

Results:
Ln 299: change "of" to "for"
Ln 309: Correct to "Our reconstructed phylogeny..." and list specifically the figure being referred to.
Ln 324: What was the bootstrap value for the ML analyses?
Ln 327-Correct to "...(Bio15) had significant support for the PNC hypothesis (Table 1), with a moderate to weak PS and with variance..."
Ln 332 Change "is" to "was"
Ln 333: Change "are" to "was"
Ln 334: Delete "are" and add "models" after "equivalent"
Ln 335: Delete "of" and add "in support of"
Ln 336-337: Correct to "...This implies weak support for the PNC hypothesis for Bio19 using the ...."
Ln 339: change "are" to "is"
Ln 340: add "of trait evolution tested." after "models"
Ln 341: change "in" to "for"
Lns330-345: My interpretation of the data in Table 2 are that a BM model cannot be excluded as the best fit model in almost all of the variables (Except Bio19). The only variable to show conclusive support for one model (according to published criterion for distinguishing trait evolution models using AICc differences >2) here is Bio15 - which shows conclusively a BM model fits relative to others. Almost all of the discussion and interpretation of other results (i.e., that a BM model does not fit) is based on these analyses - which show no definitive conclusions. This is likely due to limited sample sizes in the current analyses - but this is not acknowledged until a small comment in the discussion - there should be acknowledgment of this problem before then. Therefore, there is way too much emphasis placed on the reliability of these results and they are vastly overstated in their importance and conclusiveness throughout the ms. This is simply over-inflating confidence in the conclusions and all language related to the reliability of these tests should be modified throughout the ms given these facts.
Ln 346: Change "are" to "were"
Ln 347: Correct to "...species, and within the toquatus and poinsettii clades. Similarly, only a few species paris show.."
Ln 348: Correct to S. cyanstitctus
Ln 349: Correct to "...these were sister taxa or closely-related species in our analyses, with...
Ln 353: add an s to "layer"
Ln 354-355: Correct to "...,excepting Bio15."
Ln 358: Which variables specifically?
Ln 361: Correct to "...peaks indicate that similar climatic tolerance between a few species was observed in..." Also, which species specifically?
Ln 362: change "are" to "was"
Ln 363: delete "the" before "Precipitation", change "has" to "had"
Ln 364: Correct to "...that was narrower than other bioclimatic layers, which is consistent with an OU model of trait evolution with a single optima, as was detected...". Also, here the OU model cannot be distinguished from a model where divergence increases towards the present (as indicated by a delta >1). The delta model arguably receives support from increased MDI toward the present in DTT plots for this variable - albeit not outside the CI of the null BM model. This would contradict this statement.
Ln 366: Correct to "...S. serrifer, which shows more extreme..."
Ln 368: Correct to "...plots detailing the history of climatic tolerance evolution..." Also - what you are looking at here is not climatic tolerance evolution if you have used the differences between species as outlined in the methods. You are looking at the history of similarity and divergence of climatic tolerance. If you used the weighted means of PNO's and not their differences this statement would be correct.
Ln 371-377: These statements need to be modified given the problems with over-interpretation of the conclusiveness of analyses showing models of trait evolution.
Ln 377: Correct to "S. serrifer"
Ln 379: More influence on their distribution?
Ln 389: Bio 9 and 15 don't appear different to me at all relative to the other analyses and nothing early on falls outside the 95% CI of the null BM model (grey shading in DTT plots)
Ln 390-392: I disagree with this interpretation as variables almost never go outside the 95% CI for most variables here. The exceptions are Bio5, Bio8, Bio9, and maybe Bio19. They all show increasing divergence among subclades towards the present (correction of time axes for Fig. 6 will make this clearer). This fits with the delta values presented in Table 2 - but which cannot be distinguished relative to other models for these species.These are the interesting results from this analysis and they are almost not even mentioned here. You cannot interpret values that do not exceed the confidence limits of the null model (grey shading) just because they are greater than the average values from the null (dotted lines).
Ln 395-408 - I find this section overly speculative. It should be modified or deleted, also "relative time" here means nothing. Authors would usually correct these values to the relative time calibrations on their tree if they are available.

Discussion:

The problems highlighted above persist in the discussion, I have refrained from editing this as I feel there are substantive enough problems highlighted above that the discussion will need to be completely rewritten.


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: CLIMATIC NICHE EVOLUTION IN THE VIVIPAROUS SCELOPORUS TORQUATUS GROUP (SQUAMATA: PHRYNOSOMATIDAE)
Review round: 2
Reviewer: 1

Output:
Basic reporting: A) The English and grammar could still use a large amount of work. The paper is challenging to follow at times, especially when there are paragraphs that exceed a page and a half in length (lines 437-483).  The introduction has improved a great deal (but could still use better framing of their hypotheses) and I think most of the readability problems are found in the rest of the paper, especially the discussion, which is very difficult to follow. These areas (some pointed out below) need to be completely rewritten and condensed to their most important points.

B) Previously I suggested the authors discuss the cold-climate hypothesis more, and here they actually remove all mention of it from the main text. Oddly, the cold-climate hypothesis is mentioned several times in the abstract and that their study does not support it. The authors need to address this hypothesis in the introduction, why it is relevant to their study, and also how their study is testing this hypothesis or something different (which is not immediately clear). Additionally, a recent paper was published (below) that should be addressed here that may help the authors better add context to their study:

Ma L, Buckley LB, Huey RB, Du W‐G. A global test of the cold‐climate hypothesis for the evolution of viviparity of squamate reptiles. Global Ecol Biogeogr. 2018;27:279–289. https://doi.org/10.1111/geb.12730

C) Figures are improved and look readable now.

D) Authors should upload raw data to an online repository. They could use the center for open science (open science foundation) which is completely free, and they can provide private links for reviewers. https://osf.io
Experimental design: A) The rationale behind the study is not well explained. From what I could ascertain, the authors argue that viviparity should constrain the group to remain in cold climates because viviparity constrains other traits, so there might be some correlation in climate to these constrained traits. I do not think this is a strong argument. Viviparity presumable evolved in cold climates to deal with the lack of warmth for eggs in the external environment. However, it should be explained why this should act as a constraint in climatic variables after the trait has evolved. Are there studies that suggest that viviparity is disadvantageous in warmer climates?

B) What this study tests differently from Lambert & Wiens (2013) is not clear. Lambert & Wiens 2013 tested the cold-climate hypothesis on the same species group as this paper, and show that this species group represents an invasion of warmer climates from cooler climates, which already provides evidence against the hypothesis of PNC they aim to test here. The authors need to distinguish the two studies more clearly if they are different. If they are actually testing the same hypothesis, then this needs to be clear because PeerJ allows replication of prior studies, and different analyses are especially welcome. The true aim should not be as obscure as it is presented. The authors state their study is quite different than Lambert and Wiens 2013 but it appears quite similar, for example, Figure S6 here is basically showing the same thing as Lambert and Wiens 2013 Figure 1.
Validity of the findings: A) It is not clear how the conclusions link back to their original research question, as the authors waver between testing the cold-climate hypothesis (explicitly stated in the abstract) or whether viviparity constrains species to inhabit cold climates (stated in the introduction). If they are testing the cold-climate hypothesis, they should link this constraint back to it and why it would be a prediction of this hypothesis. If not, they should distinguish their hypothesis from the cold climate hypothesis, and explain both clearly in the introduction. I could envision arguments going both ways here.

B) More explanation for the lack of a positive result for PNC is needed. Lambert & Wiens 2013 suggest that phylogenetic history could explain retention of viviparity because viviparity evolved first in cold climates, but that cold climates are merely a prerequisite for viviparity to evolve, not a constraint as they show multiple colonizations of warmer climates by these viviparous species.

C) Why should we expect viviparity to act as a constraint for cold climates? Could this be reframed as a general expectation of PNC to cold climates? Since viviparous species occur in warmer climates, what previous research could be addressed to explain this?
Additional comments: This manuscript addresses climatic niche evolution in lizards and has improved since the last review in some ways, and has worsened in others. For this re-review, I also took into account what the other reviewers had suggested. Most of the analyses appear to be sufficient to address their hypotheses and the additional analyses are welcome, as as well as the requested changes. I have a few line item comments below:

Line item comments:

Line 43: Unclear who the “some authors” are. The cited paper goes into detail on the numerous ways to test for PNC, so it clearly not this paper you are referring to.

Lines 238: Why didn’t you test the white noise model? Support for a white noise model would support the hypothesis that there is no PNC. Otherwise the negative result from the other models could suggest many things, including a lack of power.

Lines 438-442: Lambert and Wiens 2013 does address whether viviparity constrains these species to cold-climates. They discuss in detail the paradox between so many viviparous species inhabiting warmer climates and suggest it is due to the trait first evolving in cold-climates and the species later dispersing to warmer climates.

Lines 437-483: Very long paragraph. Its also not entirely clear what the point of this paragraph is, as it just basically lists potential traits that could be related to climatic variables. I’m not entirely sure why its relevant.

Lines 498-500: This sentence is not clear.

Line 498, 500: Ravell is spelled “Revell”

Lines 541-578: Another long paragraph that is a page long.


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: CLIMATIC NICHE EVOLUTION IN THE VIVIPAROUS SCELOPORUS TORQUATUS GROUP (SQUAMATA: PHRYNOSOMATIDAE)
Review round: 3
Reviewer: 1

Output:
Basic reporting: I have reviewed the article “Climatic niche evolution in the viviparous Sceloporus torquatus group (Squamata: Phrynosomatidae)”. The study does not have methodological or sampling issues and the analyses are proper for their questions. The authors have followed most recommendations of reviewers and editor. Therefore, I would recommend this revised manuscript for publication. However, I still recommend the authors to review the English for a few mistakes I found (see some examples below in minor comments) and maybe work a little more to make a more readable and straight to the point manuscript, especially in the discussion section (see major comment below).
Experimental design: no comment
Validity of the findings: no comment
Additional comments: Major comments

The discussion is sometimes hard to follow. It tends to over discuss by comparing the results of the different analyses they performed but, in many cases, not making these discussions in terms of their predictions but in terms of how congruent are (or are not) the different approaches they used to test PNC and the theory behind this.

The authors should mention which phylogenetic tree they used for their comparative analyses. Was this the consensus? The dated tree? I couldn’t find this information in the text.

Minor comments

L7-8 It’s not clear what they meant by “using bioclimatic variables relevant to each viviparous group”. This is one of the justifications for their study but it was not clear to me, how the previous studies did not use relevant variables?

L65 the word “interactions” might be more appropriate than “variables”

L66-67 not clear what they meant by “the environmental space of non-interacting abiotic variables”

L64-68 Thus what definition of niche are they using in their study? Not clear why they discussed the differences on these definitions but not connected them with their study/questions

L115 specify that the 1km x 1km it’s an approximation

L146 “Final Maxent analysis” not sure if it should be plural

L151-252 “The identification of best-fit model” sounds rare to me

L282 “I tended to yield higher values” are they referring to their own results? Or this is a generality of this statistic?

L286 “resampling of overlap matrix” I think it should be “resampling of the overlap matrix”

L298 similar than the latter for “weighted means of PNOs”

L318 “For total phylogeny” sounds rare

L511-513 how PC3 represents dry ambient? Because the highest loadings are related to precipitation variables?

L530 “the ecological differences that detected in PNO profiles” sounds rare, maybe “the ecological differences that were detected in the PNO profiles”?

L586-589 Clarify that the bioclimatic variables used are not able to detect these microclimatic differences because of a scale problem of grain size

L640 what do they mean by “PC3 evolution”


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: CARABID COMMUNITY STRUCTURE IN NORTHERN CHINA GRASSLAND ECOSYSTEMS: EFFECTS OF LOCAL HABITAT ON SPECIES RICHNESS, SPECIES COMPOSITION AND FUNCTIONAL DIVERSITY
Review round: 1
Reviewer: 1

Output:
Basic reporting: 'no comments'
Experimental design: 'no comments'
Validity of the findings: 'no comments'
Additional comments: Dear Authors,
I read the paper with great interest.
It deals with carabid community structure and functioning in relation to grassland types.
The most original and interesting result is that carabids are strongly influenced by local climatic factors, and the responses of carabid communities to climate and other factors vary according to the grassland type.
I appreciated the analytical approach in considering the carabid community by structural and functional approach.
I added few comments directly into the pdf.
Good luck!


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: CARABID COMMUNITY STRUCTURE IN NORTHERN CHINA GRASSLAND ECOSYSTEMS: EFFECTS OF LOCAL HABITAT ON SPECIES RICHNESS, SPECIES COMPOSITION AND FUNCTIONAL DIVERSITY
Review round: 1
Reviewer: 2

Output:
Basic reporting: Technically, this submission meets the standards of PeerJ. The paper is well written and the language acceptable. The literature is reasonably cited. Some of the figures can go into an electronic appendix. Figure labels are too small and hard to read. The necessary raw data are supplied.
Experimental design: This paper reports original scientific work that meets the scope of PeerJ. However, I have reservations regarding the research questions. This is another paper that starts with global change and end in a classic faunistic study. In fact it reports niche occupancy and changes of ground beetle communities across three habitat types. Numerous studies have reported on that and this paper adds another case study. This has nothing to do with global change. Thus I cannot see the gap in knowledge filled by this paper (except the local faunistic analysis). I advise a respective rewriting of the introduction. Particularly, the starting hypotheses seem to be ad hoc. They are quite obvious and reflect what we already know. Some hypotheses rather serve as strawman for the discussion. This makes the study confirmatory and descriptive.

I have also some reservations against the methods. Pitfall samples of ground beetles are common and well introduced. However, the limitations are also known. For instance, too small pitfall sizes heavily bias the results. I expected mentioning this problem. Further, this study is a classic case of pseudorreplication. Three habitats have been studied and the pitfall data within these three habitats are not independent. Thus CCA and multiple regression results are also biased (too low standard errors). There are several ways to reduce this bias. Most often used are eigenvector maps. In the present case you might also use a generalized linear mixed model with site identity as categorical random variable or the plot distances as metric covariate. As it stands the multiple regression results might be misleading. Further, the high number of variables and comparisons open the door for false positives. This should have been corrected for.
I’m also concerned about the rarefaction approach. Rarefaction is a method of standardization of sample data. In your case you extrapolate to estimated richness. Thus I suspect you have used one of the Chao estimators. This should have been stated clearly.
Validity of the findings: Given the methodological uncertainties, the discussion is rather speculative.
The introduction highlights species composition and functional aspects. However, major part of the results and discussion is on richness only. Compositional diversity and functional aspects have not been studied sufficiently. For instance, what about beta diversity, niche overlap and variability, proportional changes in abundance, small scale heterogeneity, etc.?
The discussion only marginally reflects the global change attitude of the introduction.

Unfortunately, the conclusions are quite obvious and well documented by numerous other studies. I think this study is suited (in a revised version) for a specialized carabidological journal.
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: CARABID COMMUNITY STRUCTURE IN NORTHERN CHINA GRASSLAND ECOSYSTEMS: EFFECTS OF LOCAL HABITAT ON SPECIES RICHNESS, SPECIES COMPOSITION AND FUNCTIONAL DIVERSITY
Review round: 1
Reviewer: 3

Output:
Basic reporting: see attached document
Experimental design: see attached document
Validity of the findings: see attached document
Additional comments: see attached document


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: FLUOXETINE-INDUCED ALTERATION OF MURINE GUT MICROBIAL COMMUNITY STRUCTURE: EVIDENCE FOR A MICROBIAL ENDOCRINOLOGY-BASED MECHANISM OF ACTION RESPONSIBLE FOR FLUOXETINE-INDUCED SIDE EFFECTS
Review round: 1
Reviewer: 1

Output:
Basic reporting: A very interesting and relevant work. The sample size is large and looks very elaborate.
Experimental design: The experiments are well detailed.
Validity of the findings: Has a microbiota markers been used to compare groups as an endogenous marker?
Additional comments: Well detailed and consistent 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: FLUOXETINE-INDUCED ALTERATION OF MURINE GUT MICROBIAL COMMUNITY STRUCTURE: EVIDENCE FOR A MICROBIAL ENDOCRINOLOGY-BASED MECHANISM OF ACTION RESPONSIBLE FOR FLUOXETINE-INDUCED SIDE EFFECTS
Review round: 1
Reviewer: 2

Output:
Basic reporting: no comment
Experimental design: no comment
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: FLUOXETINE-INDUCED ALTERATION OF MURINE GUT MICROBIAL COMMUNITY STRUCTURE: EVIDENCE FOR A MICROBIAL ENDOCRINOLOGY-BASED MECHANISM OF ACTION RESPONSIBLE FOR FLUOXETINE-INDUCED SIDE EFFECTS
Review round: 1
Reviewer: 3

Output:
Basic reporting: Well written and appropriately structured manuscript.
Experimental design: Sufficient level of detail in methods and satisfactory experimental design. Concern surrounding reproducibility.  Ten per test group is quite low when looking at such variable measures as behavior and microbiota.
Validity of the findings: Concern surrounding reproducibility.  Ten per test group is quite low when looking at such variable measures as behavior and microbiota.  It is remarkable that statistical significance was achieved. Although these dramatic effects many represent a big impact, published results should be the product of 3 independent experiments.  So it is recommended that the study be repeated.
Additional comments: The manuscript is well written and clearly states aims and conclusions of study.
More ‘quantitative’ figures, outlining phyla abundances and OTUs should be included in the main figures, rather than as supplemental figures, rather than just the heat maps.  Perhaps find another way than tables to represent these data.
Figure 1: Data should be presented as mean weight ± standard deviation.  Fluoxotine vs. control should be directly compared at each time point and ANOVA applied to the data to determine whether any differences in overall weight are seen between groups.  Add n= 10 per group and appropriate figure legend, use format similar to figure 2.


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: FLUOXETINE-INDUCED ALTERATION OF MURINE GUT MICROBIAL COMMUNITY STRUCTURE: EVIDENCE FOR A MICROBIAL ENDOCRINOLOGY-BASED MECHANISM OF ACTION RESPONSIBLE FOR FLUOXETINE-INDUCED SIDE EFFECTS
Review round: 2
Reviewer: 1

Output:
Basic reporting: good
Experimental design: good
Validity of the findings: good
Additional comments: The authors have addressed all my concerns and the manuscript is now suitable for 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: FLUOXETINE-INDUCED ALTERATION OF MURINE GUT MICROBIAL COMMUNITY STRUCTURE: EVIDENCE FOR A MICROBIAL ENDOCRINOLOGY-BASED MECHANISM OF ACTION RESPONSIBLE FOR FLUOXETINE-INDUCED SIDE EFFECTS
Review round: 2
Reviewer: 2

Output:
Basic reporting: Well written.  Satisfactorily responded to all reviewer comments.
Experimental design: Appropriate
Validity of the findings: As a pilot study ok, but findings should be reproduced.
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: DESCRIPTIVE ANATOMY OF THE LARGEST KNOWN SPECIMEN OF PROTOICHTHYOSAURUS PROSTAXALIS (REPTILIA: ICHTHYOSAURIA) INCLUDING COMPUTED TOMOGRAPHY AND DIGITAL RECONSTRUCTION OF A THREE-DIMENSIONAL SKULL
Review round: 1
Reviewer: 1

Output:
Basic reporting: The study is clear, and the English used in the text is professional and very well written.
The references reported cover sufficiently the areas mentioned in the text, but I have provided a suggestion for an addition related to some background info on the diversity and extinction of the group mentioned in the paper.
A rich series of figures and tabled data are shared with the reader providing an important source for professionals interested in this group.
The structure of the article does conform to the ‘standard sections’ indicated by the Instructions for Authors PeerJ’s regulation.
The figures present in the paper are contextualized throughout the text, of excellent quality and properly labelled.
Data figured and described in the text are also provided via a 3D supplementary material file.
Experimental design: As this study represent a redescription with a new taxonomic interpretation for an historical specimen of Mesozoic marine reptile, it represents an original primary research study.
This submission defines background and aim of the study clearly, honestly and without overselling it.
The investigation performed is rigorous and combines classic biostratigraphical, anatomical observations of fossil material, and cutting edge medical imaging techniques, reporting appropriately the machinery utilized herein and information useful to replicate the study or the methodology to whomever might be interested in doing so.
Validity of the findings: See general comment below. As the study represent a morphological, taxonomic and anatomical description, it is well self-contained and doesn’t report any speculation throughout.
Additional comments: General comments for the review of Lomax et al. “Descriptive anatomy of the largest known specimen of Protoichthyosaurus prostaxalis (Reptilia: Ichthyosauria) including computed tomography and digital reconstruction of a three-dimensional skull”.

This is an excellent, well-detailed study describing a historically valuable specimen housed at the Thinktank, Birmingham Science, UK. Although the specimen was originally identified as belonging to Ichthyosaurus communis (one of the most common attribution for Jurassic ichthyosaur specimens found in Great Britain), Lomax and coauthors performed a great deconstructionist study to investigate the identity of this specimen, making it scientifically valuable and available to the community.
Lomax and colleagues traced down the original locality, constraining the stratigraphic contest and age of the specimen, described the anatomy in detail with the aid of non-invasive 3D imaging reconstruction, and reviewed its original taxonomic attribution.
Considering that many Early Jurassic ichthyosaur specimens are often preserved in 2D, this specimen represents a rare opportunity to cast some additional light into the inner soft anatomy, particularly of the skull. In this paper, the authors not only describe the osteology of this specimen, but also restore and detail the inner soft anatomy of the brain and other related structures, providing a much useful example for the anatomy of this taxon to ichthyosaur workers.
The description they present and the taxonomic attribution are convincing.
The study is very well written and acceptable as it is, but I have flagged a few comments in the review pdf (see attached file) that may improve an already worthy to be published study.
In particular:

Line 40: I would add a citation here: I’d suggest Fisher et al. 2016.

Line 85: Add size information between brackets here: although the authors mention this information further on close to the end of the paper, it is a little bit frustrating mentioning it at first and then not giving away the information. The reader will appreciate more the immediacy, which will also be found more easily by interested readers.

Line 159: remove ‘and’ and replace it with ‘like’. I know perfectly what the authors mean here, but some relatively more unaware readers may interpret this as considering dinosaurs non-tetrapods (which we know not being the case).
In conclusion, I congratulate with the authors of this contribution and recommend the study for publication in PeerJ.

Reference

Fischer V, Bardet N, Benson RBJ, Arkhangelsky MS, Friedman M. 2016 Extinction of fish-shaped marine reptiles associated with reduced evolutionary rates and global environmental volatility. Nature Communications. 7, 10825. (doi:10.1038/ncomms10825)

Alfio Alessandro Chiarenza
Imperial College London


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: DESCRIPTIVE ANATOMY OF THE LARGEST KNOWN SPECIMEN OF PROTOICHTHYOSAURUS PROSTAXALIS (REPTILIA: ICHTHYOSAURIA) INCLUDING COMPUTED TOMOGRAPHY AND DIGITAL RECONSTRUCTION OF A THREE-DIMENSIONAL SKULL
Review round: 1
Reviewer: 2

Output:
Basic reporting: Lomax and co-authors carried out CT scanning of  a large Lower Jurassic ichthyosaur and
provide very useful and needed information on the skull anatomy of this peculiar reptiles.
CT scanning provides crucial information on the anatomy of extinct forms and it is much needed in Ichthyosauria literature (up to now only 2 contributions including this). The authors explore, by means of medical computed tomography scanner, a large skull of a Mesozoic marine reptile and provide three-dimensional digital reconstruction of a large skull of an ichthyosaur. In this way they bring a reliable background for the correct interpretation of skull anatomy and highlighted the utility of medical imaging techniques as powerful tool for exploring and studying even large specimens.

Certainly is the kind of manuscript that it will become a mandatory reference for all contributions focused on systematic and diversity of Jurassic ichthyosaurs. Reliable anatomical information is the only solid framework for hosting higher levels of inferences (such as phylogenetic and palaeoecological hypotheses) so I hope that this contribution will encourage others to go on exploring the anatomy of the ichthyosaurs from other ages and also for other marine reptiles.
I recommended its publication after minor revision
Experimental design: No comment.
Validity of the findings: No comment.
Additional comments: I congratulate the authors for this contribution. The authors did an outstanding job with the CT scanning, description and reconstruction of a large 3D preserved skull of Protoichthyosaurs. The detailed information and figures provided herein will be useful and a mandatory reference for all contributions dealing with the systematic and phylogeny of Jurassic ichthyosaurs.

The paper is self-contained, well written, it has a logical flow, and it can be read without ambiguities throughout. As I am not a native speaker I don´t feel confident to review the English grammar and/or provide further comments on the English.
I made only few suggestions and comments (see annotated PDF for more particular issues).
In summary, I think it is a valuable and needed contribution to our understanding of Lower Jurassic ichthyosaurs and it provides a solid basis to house higher levels of hypotheses on the evolution of the whole lineage. I recommend its publication after minor revisions

Minor comments and suggestions:

I think that some topics are worthy to be remark and/or certain questions that could be addressed such as:

What is the significance of the new anatomical information in a phylogenetic framework? The new information permits to complete the character scoring of the taxon? This will be very useful to determine the phylogenetic location of Protoichthyosaurus.

Which is the interpretation of the peculiar vascular system of the snout? Is it similar or not to the one described by Foffa et al. 2014. in pliosaurs?

It could also be worthy if the authors can provide more details and/or reference on how the total body length is estimated.

Several localities along the United Kingdom (UK) are mentioned throughout the text and there is no map. English fossiliferous localities (particularly southern ones) are worldwide famous so some readers could be family with most of them but not with all of them. The focus of the contribution makes not necessary to include a map but, if localities are mentioned it would be necessary to explain the significance and/or meaning of these information. Maybe the locality references must not only restrict to actual political divisions of the territories but also to geological formations outcropping in more general areas like Southern, center, northern… England or Scotland, etc.

Maybe the authors could include a reference: “Sollas (1916) The skull of Ichthyosaurus, Studies in Serial Sections…”. This contribution is, as far as I know,  the first study (using different technic, of course) that provides serial sections of the skull of an ichthyosaur.

Finally, I have a minor comments on the taxonomical identification. From the two characters used by the authors to separate the two species of Protoichthyosaurus (maxilla and postorbital morphologies) I think that in the case of the BMT 1955.G35.1 only the maxilla can be used as the postorbital is lost. In any case, if the authors think that the morphology of the maxilla is not enough the specimen could be referred as Protoichthyosaurus sp.


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: DESCRIPTIVE ANATOMY OF THE LARGEST KNOWN SPECIMEN OF PROTOICHTHYOSAURUS PROSTAXALIS (REPTILIA: ICHTHYOSAURIA) INCLUDING COMPUTED TOMOGRAPHY AND DIGITAL RECONSTRUCTION OF A THREE-DIMENSIONAL SKULL
Review round: 1
Reviewer: 3

Output:
Basic reporting: The authors present a new description of an ichthyosaur specimen and assign it to the taxon _Protoichthyosaurus prostaxalis._ This is well-presented and provides a good description of this material based on CT scans that is of suitable length for the report.

I have found some minor issues that are included in this document and the accompanying marked-up PDF, and suggest **moderate revision** of the work before publication. I also think that some further comparison and discussion of the anatomy of this taxon is necessary, particularly in the context of infrequently-preserved palatal elements. I am happy to be identified and contacted further by the authors to clarify any of my comments.

The English throughout is correct and used, however, in some case the lack of proper anatomical orientation nomenclature makes the description confusing. I have marked such instances in the accompanying PDF.

The introduction and literature review are suitable and reference relevant articles. Structure is appropriate.

The figures given are not as high-quality or structured as well as I think they should be. In particular, figs 2 and 4, which are the main references for the description of the anterior skull and jaw, are too small for purpose: these _are_ the main images to point features that are mentioned in the text and thus should be much larger. Even at full screen width on my laptop, they are too small and low-resolution for me to certainly see many features that you mention. This may partially be due to the resolution of the scans, but given the JPEG(?) artefacts I do not think that is the whole case. The 3D PDF provides this information, but without the essential labelling and scaling that the static figures are there to give. Additionally, I find the colour of the premaxillae to be difficult to see on a white background; this should be changed. Perhaps also consider the colouring and its visibility for colour blind people.

The RAW data of the complete skull scans in medical CT has been included and matches the segmented images (figs 2, 4) and 3D PDF as expected, but I did not get the scan data for the μCT scans for the braincase elements. These must be made available also.
Experimental design: Research is original and well-defined, relevant, and meaningful: the new description as some detail to the knowledge of ichthyosaur cranial anatomy.

The methods used are well explained, but some more details of the CT scan settings are required for complete reproducibility: particularly exposure length, filtering used. More details are needed on how and why a similar voxel size was used for the braincase elements.
Validity of the findings: The results and discussion are well supported, but could benefit from additional comparision with other know material; this is almost entirely lacking apart from the justification for placing in _Protoichthyosaurus prostaxalis._ In particular, fuller comparison against the limited palatal and endocranial material of other ichthyosaurs will be useful as a marker of ichthyosaur evolution, and morphological variation. Examples where some the palate is known include _Ichthyosaurus, Ophthalmosaurus, Platypterygius, Stenopterygius, Sveltonectes_ [McGowan 1973; Moon 2016; Baur 1895; Kear 2005; Fischer2011], which would all make pertinent comparative material.
Additional comments: # Additional considerations #

A few particular language considerations:

* Use anatomical orientations where required: e.g. 'lateral' or 'medial' (as relevant) over 'right' or 'left' (l 261); 'laterally' rather than 'outwards' (l 365).
* Ensure that directions 'left' or 'right', where used, are used correctly: in some instances it was unclear whether you were referring to left or right or paired elements, or to the left or right side of an element (e.g. description of the premaxilla \[l 261\]). More often these should be replaced with 'medial' and 'lateral'.

Several elements are described or mentioned as being present, but not given a formal description:

* Right nasal (l 299): mentioned as present but not figured.
* Postfrontal (l 323): mentioned as present and partially complete, but neither described nor figured.
* Supratemporal (l 425): additional figures of the lateral margin to show the additional facets should be included.

These elements should be figured to show they are difficult to describe, or should be included more fully.

## Maxilla ##

Does the maxilla show evidence of the external narial margin on its dorsal surface? This is one of the features of _P. prostaxalis_ described by @Lomax2018, as it would be useful to compare.

## Jugal ##

The left side, as preserved, shows this it extends anterior to the orbital margin, as marked by the prefrontal, but does not appear t be complete. Additionally, the facets on the lacrimal and maxilla appear to extend anterior to this. Is the extent of the left jugal going to go extensively anterior to the anterior orbital margin or minimal?

## Pterygoid ##

The comparisons between BMT 1955.G35.1, _Sveltonectes,_ and _Ophthalmosaurus_ (l 390) may need more justification: what are the similarities/differences? bearing in mind that the pterygoids of both this specimen and _Sveltonectes_ are incomplete compared to _Ophthalmosaurus._ The general form seems to be similar in the shape of the quadrate ramus, while only the posterior portion of the palatal ramus is preserved in BMT 1955.G35.1.

I can't see the dorsal ramus of the pterygoid clearly in either the figures or the 3D PDF. Can this be pointed out somewhere? Also, there is no description of the contact laterally with the quadrate: how extensive? how close? does it meet the stapes (as in _Ophthalmosaurus_ and _Leninia_ [Fischer 2014, Moon 2016])? These details, or mention that it is not describe-able.

## Quadrate ##

The groove dividing the jaw articulation into two sections is common in ichthyosaurs, so I don't know why you have picked out only _Acamptonectes._ Other mentions include: _Ichthyosaurus_ [McGowan 1973], _Ophthalmosaurus_ [Moon 2016], _Platypterygius_ [Kear 2005].

# References #

Baur, G. 1895. “Die Palatingegend der Ichthyosauria.” Anatomischer Anzeiger 10 (15): 456–59.

Fischer, Valentin, Maxim S Arkhangelsky, Gleb N Uspensky, Ilya M Stenshin, and Pascal Godefroit. 2014. “A New Lower Cretaceous Ichthyosaur from Russia Reveals Skull Shape Conservatism Within Ophthalmosaurinae.” Geological Magazine 151 (01): 60–70. doi:10.1017/S0016756812000994.

Fischer, Valentin, Edwige Masure, Maxim S Arkhangelsky, and Pascal Godefroit. 2011. “A New Barremian (Early Cretaceous) Ichthyosaur from Western Russia.” Journal of Vertebrate Paleontology 31 (5): 1010–25. doi:10.1080/02724634.2011.595464.

Kear, Benjamin P. 2005. “Cranial Morphology of Platypterygius Longmani Wade, 1990 (Reptilia: Ichthyosauria) from the Lower Cretaceous of Australia.” Zoological Journal of the Linnean Society 145 (November): 583–622. doi:10.1111/j.1096-3642.2005.00199.x.

Lomax, Dean R, and Judy A Massare. 2018. “A Second Specimen of Protoichthyosaurus Applebyi (Reptilia: Ichthyosauria) and Additional Information on the Genus and Species.” Paludicola 11 (4): 164–78.

McGowan, Christopher. 1973. “The Cranial Morphology of the Lower Liassic Latipinnate Ichthyosaurs of England.” Bulletin of the British Museum (Natural History), Geology 24 (1): 1–109.

Moon, Benjamin Christopher, and Angela M Kirton. 2016. “Ichthyosaurs of the British Middle and Upper Jurassic. Part 1, Ophthalmosaurus.” Monograph of the Palaeontographical Society 170 (647): 1–84. doi:10.1080/02693445.2016.11963958.


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: DESCRIPTIVE ANATOMY OF THE LARGEST KNOWN SPECIMEN OF PROTOICHTHYOSAURUS PROSTAXALIS (REPTILIA: ICHTHYOSAURIA) INCLUDING COMPUTED TOMOGRAPHY AND DIGITAL RECONSTRUCTION OF A THREE-DIMENSIONAL SKULL
Review round: 2
Reviewer: 1

Output:
Basic reporting: I have no further comments to those expressed in my previous review. I agree with the answers and / or refutations to my former comments. I congratulate the authors for this interesting contribution that will become a mandatory reference for future research on Jurassic ichthyosaurs. I hope that this contribution encourages others to deepen the anatomical exploration of these great marine reptiles.
Experimental design: no comment
Validity of the findings: no comment
Additional comments: I have no further comments to those expressed in my previous review. I agree with the answers and / or refutations to my previous comments. I congratulate the authors for this interesting contribution that will become a mandatory reference for future research on Jurassic ichthyosaurs. I hope that this contribution encourages others to deepen the anatomical exploration of these great marine reptiles.


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: DESCRIPTIVE ANATOMY OF THE LARGEST KNOWN SPECIMEN OF PROTOICHTHYOSAURUS PROSTAXALIS (REPTILIA: ICHTHYOSAURIA) INCLUDING COMPUTED TOMOGRAPHY AND DIGITAL RECONSTRUCTION OF A THREE-DIMENSIONAL SKULL
Review round: 2
Reviewer: 2

Output:
Basic reporting: I find this to be appropriate and thorough throughout. My comments from the first version have been addressed suitably.
Experimental design: As the original scan data has been lost, I think this should be mentioned either in-text or in the supplement to explain why it is not included; this will (hopefully) prevent the authors being asked in future why it wasn't included too.
Validity of the findings: No comment
Additional comments: Thank you for your replies to the modifications I requested.

A lot of my comments concerned presentation of figures, so I wish to justify myself: my view is that as much of what is discussed in text should be included in the figures alongside as possible, particularly described features, and even if a separate 3D or other data set is available. I realise this is not always possible, but have tried to match these standards. This is why I requested larger, higher resolution images, and more images of elements that did not show the described features. It is possible in Avizo to export images larger than the window size using the 'Render tiles' option, and avoiding JPEG compression where possible by using TIFF of vector files is a good option. The supratemporal in Fig 6A–C does not clearly show all the facets described.

However, given the presence of the 3D PDF I am happy with the quality and completeness of the figures presented by the authors if the editors are also.


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 EFFECT OF CYCLING HYPOXIA ON MCF-7 CANCER STEM CELLS AND THE IMPACT OF THEIR MICROENVIRONMENT ON ANGIOGENESIS USING HUMAN UMBILICAL VEIN ENDOTHELIAL CELLS (HUVECS) AS A MODEL
Review round: 1
Reviewer: 1

Output:
Basic reporting: The work described in the manuscript is clear, nevertheless I would recommend a revision of the English used. Especially, I would suggest to carefully revise the punctuation, as it affects the understanding of the message and also the flow of the reading.
In many cases a ";" is used in the place of "," and several subordinates are not in agremment with the main sentence.

I have found an appropriate descrition of the background with recent literature references.

I would recommend to improve the quality of the figures and of the figure legends.

In my opinion, although the results are ineteresting, the authors need to explain better and elaborate the role of the different treatments the experimentally reproduce, in the light of their finding and give a link of these "arificially induced" hypoxia to the physiological situation that may occur in tissues.
Experimental design: Experiments are clear enoghu and the methods are reported in details.
Validity of the findings: Some conclusions are drawn without a clear and robust evaluation (e.g. the role of VEGF in changing cell shape an migration ability).
I would require a better description of the mammosphere generated, especially giving some  quantifiable paramenters, not only a picture to show the appearence of the cells.
Additional comments: The authors make clear which is the purpose of their study at the end of the introduction, following a logical scheme. Nevertheless, they do not give any background about the drug they intend to use (doxorubicin). I suggest to briefly explain the effect of the drug and maybe give a reference.
The workflow shown in Figure 1 is unnecessary, as the experiment flow has been stated at the end of the introduction. To my opinion, the graph does not anything to the description.
Figure 2: it is evident that between day 2 and day 7 the clumps and clusters appear bigger, but is there a positive control to show? Or a specific staining giving evidence that these clumps are actually formed of vital cells? It is also required to label panels with A, B, C etc. as the authors refer to letters in the figure legend. The figure legend should be composed of a title and an explanation that is not the mere copy of the title. Moreover, in the results section (page 16 line 400) the authors state that after three weeks the MEF increased. Can they show a picture to highlight the final material?
Figure 3: I understand it is only a morphological observation, but the authors should give an explanation of the difference they see among the different treatments. There are no parameters or other evaluation of the mammospheres in fig. 3 A-G. What is relevant? How the intermittent shots are different from continuous in term of results?
Table 1: how do the authors explain the difference in CD44+/CD24- expression between INTR.20 and INTR.40 and between CONT.5 and CONT.15.
Table 2: how do the authors explain the difference in drug resistance observed among the different treatments? (Both CONT. and INTR.). Is there any relationship between the drug response and the level of CD44+/CD24- expression reported in the previous experiment?
Figure 8: if possible, add a positive control for the transwell migration (a purified chemokine) to give an estimation of the migration ability.
Page 20, line 571: which is the parameter determined? Cell migration or cell proliferation?
Figure 9 and 10: poor quality. Please improve the microscope images or, if not possible, show as example only a control and a treated, plus the quantification. In Figure 9 it is not possible to distinguish the migrated cells and the empty space.
The authors quantify the presence of the secreted VEGF in the medium of mammospheres and then concluded that this is responsible of the endothelial net-like structures. Although VEGF is surely a good candidate for the induction of the described ability, a direct evidence in the experiments presented is lacking. I suggest at least to add a treatment with a neutralizing specific VEGF antibody to verify the contribution of the latter in the mechanisms observed.
The authors should also give an explanation of the rationale underlining the use of different hypoxia treatments (continuous and intermittent) and discuss the findings they obtained comparing the two and within each one, the different dose-response they obtained.


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 EFFECT OF CYCLING HYPOXIA ON MCF-7 CANCER STEM CELLS AND THE IMPACT OF THEIR MICROENVIRONMENT ON ANGIOGENESIS USING HUMAN UMBILICAL VEIN ENDOTHELIAL CELLS (HUVECS) AS A MODEL
Review round: 1
Reviewer: 2

Output:
Basic reporting: This article meets the standard for publications.

1) The English used in the articles needs to proofread and edited by a language editor. At multiple places, the sentence and grammar used to make it difficult to understand.

2)The literature references and background context have been provided sufficiently.

3) Figure and tables have been structured appropriately.

4) hypothesis in the manuscript have been supported by relevant results.
Experimental design: The research presented in this manuscript is within the general scope of PeerJ journal. Overall the methods have explained, and the experiments have been designed to answer the question asked at each stage of the manuscript. Sufficient "n" has been used and the correct statistical test has been performed.
Validity of the findings: Authors use AnaeroGen Compact to create a hypoxic environment and make an assertion that it creates an environment of less than 1% oxygen. Have the authors confirmed then with some electronic or biochemical test? If not they should mention that AnaeroGen was used to create a Hypoxic environment.

For the INTR and CONT treatments used and motioned in the paper. How did the authors arrive at this conditions, what is the rationale for these specific conditions was it and trial and error method or was it based on from previous publications.

Authors need to show some evidence that during INTR or CONT incubation periods cells do experience the hypoxic condition. One possible way would be to Western blot for HIF immediately after hypoxia exposure. (The authors would have to be quick for collecting the cells has HIF1a has short half-life)

Authors use CD44+ positive bead selection. This could be a point of concern as in multiple cases the beads get bound to cells for a prolonged period of time and lead to stimulation of cells in an unintended way. Flow showing Enrichment of cd44+/cd25- after magnetic bead sort should be shown in the supplemental figure.

Migration of HUVEC and Elisa show that under hypoxia Cbc cells have a secretory phenotype. It would be informative to profile the cytokines in the media using multiplex bead-based assay like luminex.

Do authors think that these CSC between INT and CONT are differnet metabolically and doing assay like oxygen consumption, glucose metabolism could sheed light on further differnence.

Minor commets:-

In mammosphere culture protocol at line 146 authors mention that 2ml of media was topped off every 2 days till day seven. In that case, the total volume would be 16 ml for in a well volume of 16.8ml which could affect gas exchange. Was the approach used as authors think that mammosphere secrete factors essential for there growth and removing the media completely would hamper the growth?

At multiple places reference to figure/table is missing.

The images are missing scale bar and have and have 3D elevation in Figure 2 and 3. Please remove elevation its distracting. Also, mention the magnification at which images were taken.
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: THE EFFECT OF CYCLING HYPOXIA ON MCF-7 CANCER STEM CELLS AND THE IMPACT OF THEIR MICROENVIRONMENT ON ANGIOGENESIS USING HUMAN UMBILICAL VEIN ENDOTHELIAL CELLS (HUVECS) AS A MODEL
Review round: 1
Reviewer: 3

Output:
Basic reporting: The manuscript is written in clear and professional English language.
The introduction is comprehensively contextualizing the issues of the subject. The objectives of the work could be better defined at the end of the introduction.
Results- numeric data are showed inside parenthesis, for example: line 428 “ In contrast, the parental MCF-7 cells had only (1 ± 0.1%) CD44+ /CD24- expression and the unsorted mammospheres had (33.2 ± 430 3%) CD44+ /CD24- expression “ . It should be: “In contrast, the parental MCF-7 cells 429 had only 1 (± 0.1%) CD44+ /CD24- expression and the unsorted mammospheres had 33.2 (± 430 3%) CD44+ /CD24- expression.
Figures- The quality of the figures are in general good. I don’t like the 3D effects in figures 2 and 3. Please, improve the quality of pictures in figures 10 and 11.
 Discussion- The first part of Discussion is repetitive of Introduction. CSC is well described in the introduction. In my opinion, it could begin in line 599.
Experimental design: The experimental approach is quite interesting and the obtained data are relevant to the better understanding of tumor cell biology. Figure 1 illustrates very well the experimental schedule.
Validity of the findings: Data are relevant and well discussed until CONTR10 and INTR30, when they observed increased in drug resistance, as well as the enhanced in capillary tubular structure formation.  The effects of INTR30 and CONT10 are not adequately discussed.
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: THE EFFECT OF CYCLING HYPOXIA ON MCF-7 CANCER STEM CELLS AND THE IMPACT OF THEIR MICROENVIRONMENT ON ANGIOGENESIS USING HUMAN UMBILICAL VEIN ENDOTHELIAL CELLS (HUVECS) AS A MODEL
Review round: 1
Reviewer: 4

Output:
Basic reporting: Basic reporting is redundant. Lacks the flow and transition, too lengthy.
Experimental design: Experimental designs are simple and easy to follow
Validity of the findings: no comment
Additional comments: 1.	The authors may want to discuss what is known about MCF7 phenotypes in 3D culture or hypoxia in general.
2.	The results and discussion are extremely wordy and sometimes deviating from the objective of this paper. Figure 2: A, B, C and D legends are missing from the figure.
3.	Figure 3: magnification of panel E and G do not look equivalent to A,B,C,D, F and H.
4.	Figure 4: Flow cytometers  dot plots. Gating for A,G and H have been moved. Needs to be exactly similar to the rest of the dot plots.
5.	For better understanding of the figure, the % population of CD24-CD44+ should be written on the dot plot itself.
6.	Figure 10 and 11: Poor representation of the images, dotted lines in the cell monolayer gap can make the images more clear.


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 EFFECT OF CYCLING HYPOXIA ON MCF-7 CANCER STEM CELLS AND THE IMPACT OF THEIR MICROENVIRONMENT ON ANGIOGENESIS USING HUMAN UMBILICAL VEIN ENDOTHELIAL CELLS (HUVECS) AS A MODEL
Review round: 2
Reviewer: 1

Output:
Basic reporting: Although the quality of English has improved, I would recommend an extra check by a mother tongue, as several mispellings are still present.
Experimental design: The experiments are well designed and the results have been presented with the higher rate of details allowed by the authors' working system.
Validity of the findings: The discussion section has been improved with indications about the different hypoxia treatments.
Figure legends have been modified.
Additional comments: The quality of the manuscript has improved. I would like to thank the authors for the detailed response to my questions. Nevertheless, there are still few issues concerning punctuation and language that need to be addressed. Here are some points:
Page 2 line 51: “…throughout the body; including…” change “;” with “,”
Page 2 line 62: “The exact signaling pathways by which hypoxia activates its effects is complex..” change “is” with “are”
Page 2 line 65: change “&” with “and”
Page 3 line 75: “chronic hypoxic exposure is incubating cells in hypoxia up to several weeks”, change “is” with “is obtained by”
Page 3 line 89: “CSCs secretions help recruit endothelial..”, change in “…help to recruit…”
Page 11 line 382: “…but their number became sufficient only 3 days after which the cells were identified via FACS analysis…” this sentence is not completely clear. Three days after what? Sorting?
Page 12 line 419: “chemoresistance; MTT assay”, change “;” with “,”
Page 13 line 433: “…that mammosphere cultures are enriched with CSCs contributing to the higher drug resistance toward doxorubicin when compared to the parental MCF-7.” Please check the grammar of this sentence.
Page 13 line 461-462: “angiogenesis in breast cancer; quantitative assays for the main angiogenic factor; VEGF-A was performed” change “;” with “,”
Page 14 line 484-486: “These findings suggest that CSCsHYP secretionscomprised in CdM from CONT.5 and INTR.20 episodes significantly promot the migration of HUVECs.” Check this sentence for grammar and meaning.
Page 14 line 493: “To clarify the interaction between endothelial cells and CSCsHYP in tumor microenvironment; HUVECs were examined”, change “;” with “,”


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 EFFECT OF CYCLING HYPOXIA ON MCF-7 CANCER STEM CELLS AND THE IMPACT OF THEIR MICROENVIRONMENT ON ANGIOGENESIS USING HUMAN UMBILICAL VEIN ENDOTHELIAL CELLS (HUVECS) AS A MODEL
Review round: 2
Reviewer: 2

Output:
Basic reporting: The article has been revised and the current version is improved in terms of english and context.
Experimental design: Experiments are clear and the methods are reported in details.
Validity of the findings: Authors have addresed all the corncerns and the manuscript is now well crafted and acceptable.
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: SIMULTANEOUS RAPID DETECTION OF HANTAAN VIRUS AND SEOUL VIRUS USING RT-LAMP IN RATS
Review round: 1
Reviewer: 1

Output:
Basic reporting: The study proposed by Zhang et al entitled: Simultaneous rapid detection of Hantaan virus and Seoul virus using RT-LAMP in rats, is in line with the state-of-art in the literature since is focused on the development of an isothermal amplification system for the diagnosis. However the novelty of the assay is limited since other report with the same aim has been previously published (Hu et al.2015. J Virol Methods. 2015 Sep 1;221:68-73. doi: 10.1016/j.jviromet.2015.04.017.), In this regards I have found some critical point that I think the authors must address before to accept the manuscript.
Experimental design: Following the direction of the OIE manual and WHO manual for the validation of diagnostic assays based on amplification of nucleic acid the analytical sensitivity of the assay to be developed needs to be assessed in the same clinical samples where assay is expected to detect the agent. For this reason, I strongly recommend to the author that they must show the sensitivity of the new assay in clinical samples and not in distilled water, since this approach is not correct.
Validity of the findings: Some critical point are missing such as:
1-Analytical sensitivity determined on clinical material where the assay is addressed (not in distilled water)
2-Analytical specificity showing the assay can amplify hantavirus regardless the gentic diversity (representative member from the main four clades)
3-A analytical comparison between the new assay and the previously reported by Hu et al.2015. J Virol Methods. 2015 Sep 1;221:68-73. doi: 10.1016/j.jviromet.2015.04.017.
Additional comments: The study proposed by Zhang et al entitled: Simultaneous rapid detection of Hantaan virus and Seoul virus using RT-LAMP in rats, is in line with the state-of-art in the literature since is focused on the development of an isothermal amplification system for the diagnosis. However the novelty of the assay is limited since other report with the same aim has been previously published, In this regards I have found some critical point that I think the authors must address before to accept the manuscript.
Major comments:
1-Another RT-LAMP for simultaneous detection of Hanta virus and seoul virus has been published in 2015, an experimental comparison between both assays showing advantages and disadvantages of the new proposed assay is needed. This comparison will allow to the reader to understand why a new assay based on the same principle that the previously reported is needed in the scientific literature.
2-Te authors designed the primers based on GQ279392.1 GenBank strains however considering the genetic diversity of Hantavirus members four distinctive clades are described. Based on a vast number of reports about RT-LAMP assays, is well know that the hybridization of B3 and BIP primers are critical for the isothermal amplification. Therefore in order to guarantee that the assay really amplify these other clades of hantavirus the author must follow two directions: i) show at least in supplementary information with an alignment of sequences including sequences from the diverse clades of Hantavirus members others than the used for the primers design, ii)use in the analytical specificity of the assays representative strains from these clades, to guarantee that not only in silico but in vitro the assay can specifically amplify diverse genetic background into the Hantavirus.
3-Following the direction of the OIE manual and WHO manual for the validation of diagnostic assays based on amplification of nucleic acid the analytical sensitivity of the assay to be developed needs to be assessed in the same clinical samples where assay is expected to detect the agent. For this reason, I strongly recommend to the author that they must show the sensitivity of the new assay in clinical samples and not in distilled water, since this approach is not correct.
Considering the elements provided above I am willing to review a new version of the manuscript with the needed amends, however in the current form I can not accept the manuscript for its 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: SIMULTANEOUS RAPID DETECTION OF HANTAAN VIRUS AND SEOUL VIRUS USING RT-LAMP IN RATS
Review round: 1
Reviewer: 2

Output:
Basic reporting: The paper is written in generally correct language, but somewhat confusing, with figures being called out of numerical order in the text, and some important issues are touched all too briefly, and should deserve more attention for the sake of clarity.

Literature review has important gaps. It is important that authors consider and discuss results previously published by others, based on studies that were conducted in the same country. For example, a very careful study was published by Hu D et al. in 2015, on the "Development of reverse transcription loop-mediated isothermal amplification assays to detect Hantaan virus and Seoul virus" (Journal of Virological Methods 221: 68–73, 2015). The lack of mention to that study is suggestive of carelessness in the literature review. The sensitivity of that assay was 10 copies (similar to PCR) for HTNV, and 100 copies for SEOV (ten-fold less than qPCR). These comparisons deserve mention.

Legends for figures are poor and some are impossible to understand entirely for lack of explanation.
Experimental design: Methods are insufficiently described.
Validity of the findings: The information is important, but it is still unclear how it relates to previous very similar studies.
Additional comments: This paper addresses the very important issue of developing sensitive and specific methods for the direct detection of the Old World hantaviruses Hantaan and Seoul in rats. However, the authors failed to make clear the reasons why their test is better than others, and a more accurate focus on that issue is needed.
Certain other issues deserve clarification.
Major concerns:
1-What hand-foot-and-mouth causing virus was used? Coxsackie? What type? (line 99)
2-How were the lungs of infected rats obtained? It is not enough to mention that they came to the authors from the Center for Disease Control and Prevention, but how were they obtained, preserved, and handled.
3-Line 210: text reads: "....in addiiton to isothermy." Sentence is too telegraphic, and it should explain why is that advantageous.
4-Figure 3 is called after figure 4, and has no explicative legend. Nobody knows what is in the tubes, and on the gel of panels B and C, respectively.
5-Lines 186-187 and Figure 4: The text says "selected as the optimal condition because of the higher specificity and enzyme inhibition at higher temperatures", but it is unclear how that was based on the curves n figure 4, since the 65 degrees curve looks almost the sabe as 66, for example. Some curves looked to be almost the same.
6-Figure 2 requires more clear legend to clarify that the tubes on panel B correspond to the viruses in the lagend of panel A.
7-Legend of figure 5 is utterly improper and unclear. Which positive samples in the curves correspond to the tubes? The comment about disagreement between tests should appear in the Results text, and not thrown in the legend of figure 5.
8-A comment should be made on the cost of this assay as compared to PCR.

Minor issues:
Line 70: the word 'is' is repeated twice.
Table 1, Line 1: Primers for the S gene.
Line 136: measured at 6-s intervals.
Line 160: I presume it should read "positive control plasmid pGEM-S was used along with every tested batch".
Line 182: HS-72 were selected as optimal primers.


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: ADVANCES IN CULTIVATION, WASTEWATER TREATMENT APPLICATION, BIOACTIVE COMPONENTS OF CAULERPA LENTILLIFERA AND THEIR BIOTECHNOLOGICAL APPLICATIONS
Review round: 1
Reviewer: 1

Output:
Basic reporting: The pictures in Figure 2 are low in resolution and are hard to see clearly. It would be better if pictures with higher qualities can be provided.

In the last senctence of the abstract, it is recommended to replace "various aspects" with the specific points that have been addressed in the paper.
Experimental design: The review is composed of several sections. In each section, one aspect of Caulerpa lentillifera is adequately discussed. One apparent deficiency is that the author did not include smooth transitions between different topics and show his or her understanding of the correlation between these different aspects. The review reads more like a list of facts, rather than a coherently organized paper. It is highly recommended that the author add more transitions between the facts to show the readers why and how the field is developed graduallly.
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: ADVANCES IN CULTIVATION, WASTEWATER TREATMENT APPLICATION, BIOACTIVE COMPONENTS OF CAULERPA LENTILLIFERA AND THEIR BIOTECHNOLOGICAL APPLICATIONS
Review round: 1
Reviewer: 2

Output:
Basic reporting: n/a
Experimental design: n/a
Validity of the findings: n/a
Additional comments: n/a


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: ADVANCES IN CULTIVATION, WASTEWATER TREATMENT APPLICATION, BIOACTIVE COMPONENTS OF CAULERPA LENTILLIFERA AND THEIR BIOTECHNOLOGICAL APPLICATIONS
Review round: 1
Reviewer: 3

Output:
Basic reporting: 1-	Literature was only focused on Caulerpa lentillifera and its future trends. Only one article was mentioned as a study of C.lentillifera, but it was a study of another alga.
Line 114 This study described the production of rhizoids in Caulerpa prolifera, not of C.lentillifera.

2-	Authors show a photo of the alga, that is adequate. However, they also considered a picture of another study and a chemical structure of a compound mentioned. They do not specify if it is a specific chemical structure detected in C.lentillifera.  Moreover, tables showing studies with specific data and the most important conclusions are necessary to facilitate the comprehension of the abilities and future trends of C.lentillifera. Tables should emphasize the characteristics of C.lentillifera and its use in different bioprocesses, such as operational conditions and their effect in growth alga,

3-	The introduction is clear and concise, but it is necessary to detail the importance of the topics discussed in this review.
Line 62-63 It is necessary to emphasize the importance of the optimum operational conditions (or cultivation conditions) to improve the productivity of the mass culturing of C.lentillifera to supply the current demands.
Line 64-70 I suggest the authors should write one paragraph describing the applications in the industry of this alga. Currently, they are focused in the treatment of wastewaters (specific compounds and the use in aquaculture) and the use of bioactive components.
4-	In general, the manuscript is written in professional English, excepting for some sentences. I suggest an English editing by native speakers.
Experimental design: 1-	Article content is adequate to this journal; however, it is necessary to describe in more detail some studies mentioned. Authors discuss different data of several studies; however, those differences can be for the operational conditions used in them. In addition, the origin of the alga should be considered to discuss the differences, because they could have a different genetical background that can influence the results.

Line 23 This alga also presents saturated fatty acids. It has been described that C.lentillifera contains a high level of unsaturated fatty acids, I suggest to specify this information in the abstract.
Line 26 These are operational conditions (or cultivation conditions) that can produce a stress in specific situations and have been studied in C. lentillifera. In addition, there are other compounds that can produce stress, such as heavy metals.
Line 27 The name species must be written with two words, genera and species, in the first time that is mentioned. After that, the name species can be mentioned as C.lentillifera, in the case of being mentioned again in this review.
Line 81 To understand the contribution of Paul et al., 2014, would be necessary to add more details of the study. Special emphasis on the operational conditions and the origin of alga that possess direct influence in the mass productivity.  It is necessary to explain the meaning of "cultivation periods".
Line 82 These are operational conditions or cultivation conditions, characteristics of the environmental. They are stress factors when their values change and produce problems in the physiology of the alga.
Line 89 It is necessary to consider the cultivation conditions in the studies mentioned, as well as the origin of the alga. It is also important to emphasize if seawater or synthetic seawater were used in these studies.
Line 96 It is necessary to know the cultivation conditions that could influence in the differences in specific growth rate in the studies mentioned. These differences can also be related to the origin of the alga
Line 97 Nitrogen possesses different oxidation states more than types of N.
Line 100 To compare the values of the salt concentrations and the growth rate of alga will give more details about their influence on the growth.
Line 102 To compare different ratios is necessary to detail cultivation conditions.
Line 103- 110 It is not clear if phytohormones were incorporated into the assays, or they were hormones of the alga.
Line 111 I suppose the authors wanted to write "cellular enzymes".
Line 117 To describe the cultivation conditions. Guo et al. (2015) performed assays at laboratory scale.  They also discuss the effect of different irradiances and other physiological data that could be included in this review.
Line 118- 125 Wu et al. (2017) also discuss other light conditions that could be mentioned as stress factors, as well as the physiological response of the alga.
Line 134 To emphasize the removal efficiency, the values and the conditions of the experiment would support the ability of the dried alga to remove metals.
Line 135 To emphasize the sorption time of the metals, to compare with other studies would support the abilities of this alga.
Line 142 As previously mentioned, to emphasize different aspects of the process, it is necessary to compare with the results of other studies.
Line 146 To know the kinetic model is adequate to understand the process. However, to include the equations are part of other topics and they are not specific of C. lentifillera.
Line 164- 165 To include information of the scaling up of the system would be interesting of considering, since the topic is the use of C.lentillifera in the wastewater treatment.
Line 167- 168 This is a characteristic of the algae, not only of C.lentillifera.
Line 176 To know the removal is better to understand the process, as well as the operational conditions.
Line 180 It is not clear if the effect is in all dyes or in specific cases.
Line 181 To know values would improve the sentence.
Line 185 Salinity is an operational condition mentioned previously, to mention the values detected is better to understand the effect in the sorption capacity.
Line 200 In the wastewater treatment the operational conditions of the process and the removal efficiency are important results that the authors should add in the discussion.
Line 215 To compare values would add information to emphasize the ability of C.lentillifera, a well as details of the assay.
Line 217 It is necessary to know the model used in this study.
Line 220 To specify the experimental conditions and the model used in this study.
Line 232 To consider the differences in the origin of the alga and the cultivation conditions of both studies could give information about the differences in the polysaccharides.
Line 250 These values were described in Codium fragile, Caulerpa lentifillera and Umbraulva japonica. It is not a number detected in several algae.
Line 267 They are operational or cultivation conditions.
Line 271- 272 To mention if assays were in cell culture or animals.
Line 284 To incorporate its use in aquaculture.
Line 287- 288 To mention if they were cell culture or animals.
Validity of the findings: 1-	Conclusions are according to the information discussed. The information requested would lead to robust conclusions.
2-	Argument set out in the introduction was developed. As mentioned above, more details of the studies are requested.
3-	The anticoagulant property of crude extract (PMC5604272), the effect of the desiccation (DOI: 10.1007/s10811-018-1442-1) and the
        immunostimulatory activity of novel polysaccharides (DOI:10.1016/j.ijbiomac.2017.12.016) are aspects of the C.lentillifera were not mentioned in this review and they have been described in the literature.
4-	Conclusions should consider the use at industry scale of the wastewater and animal test of the compounds, as future trends of the use of C.lentillifera.
Additional comments: 1-The title of the review should indicate the general applications of C.lentillifera, such as "Advances in cultivation and future trends of C. lentillifera in industrial applications (or biotechnological applications)".
2-Algae have been used in other applications, such as the production of biofuel or their use in CO2 fixation that have not been exploited in C. lentillifera and could be incorporated in the future trends of this alga.


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: ADVANCES IN CULTIVATION, WASTEWATER TREATMENT APPLICATION, BIOACTIVE COMPONENTS OF CAULERPA LENTILLIFERA AND THEIR BIOTECHNOLOGICAL APPLICATIONS
Review round: 2
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
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: ADVANCES IN CULTIVATION, WASTEWATER TREATMENT APPLICATION, BIOACTIVE COMPONENTS OF CAULERPA LENTILLIFERA AND THEIR BIOTECHNOLOGICAL APPLICATIONS
Review round: 2
Reviewer: 2

Output:
Basic reporting: The manuscript was improved, however, I have other comments that could help in the discussion. Authors should revise the measurement units. For example; in line 107 the wrote 0.6 mmol /L, and in line 136 they wrote 40 µmol photons m -2 s-1. They should write the unit according to PeerJ format. It is necessary to consider a space between number and measurement units, for example, line 111 light of 40umol.

Line 37-39 The topics; cultivation, wastewater treatment, and biological active components were previously mentioned in line 24-26. I suggest to delete them in the lines 37-39 and to use the sentence as a short conclusion.
Line 67  It is not clear if this aspect is concerned of the alga or general studies. I suggest writing the sentence again.
Line 91 I suggest changing the word "changed" to the word "change".
Line 94-98 Authors incorporated the origin of the alga, but the sentences are not clear. I write these lines again, taking care of the correct word order in the sentences.
Line 100 What groups?
Line 104 It should be written a conclusion; therefore, studies suggest that the optimal salinity concentration is between...
Line 109 Authors mention the origin of the alga, however, I suggest revising the correct word order.
Line 121 It would be adequate to write a conclusion, such as to mention the best concentration of the nutrients mentioned.
Line 124 Result of what? Results of Tao et al., 2017? It is necessary to detail it.
Line 129 I would delete the word "obviously". It is not obvious the increase of the intracellular crude polysaccharide content, this must be detected.
Line 134 Productivity of biomass?
               Authors used the word "decreased" twice in the same sentence, change for a synonym o re-write the sentence.
Line 138 The name of the gene should be in cursive.
Line 138- 139 Authors used the word "different" twice in the same sentence, change for a synonym o re-write the sentence
Line 149 This paragraph should contain some conclusions of the results discussed previously and to include future trends in the cultivation of the alga.
Line 172 It is necessary to write the reference where authors read the information.
Line 215  To write values would help to comprehend the importance of this aspect.
Line 245 I suggest writing: "as a method of wastewater methods".
Line 268 Eucheuma cottonii should be written in cursive.
Line 270- 271 the meaning of the adjectives "strong" and "weak" for the noun “scavenging activities” could be explained in a better way, for example, writing values or other studies to support the information.
Line 289 These results were inconsistent or these results were not consistent.
Line 332 Caulerpa lentillifera should be written in cursive.
Experimental design: The manuscript is adequate. I have some suggestions.
Line 92. To describe some effects in the physiology of the alga would be adequate.
Line 95 It is not clear if the analysis were done at the laboratory or industrial scale.
Line 105-106 All nutrients are important. It is better to write that they are necessary for the growing of the alga and must be taken from the environmental.
Line 165 To mention other materials and the time that they need to take heavy metals will improve the discussion.
Line 325 Information about the role of these genes could help to understand the importance of siphonaxanthin.
Line 328 It is not clear if the inhibition was in cell culture or animals.
Validity of the findings: Line 335- 337 To conclude this information, authors should include information on the topics that have not been studied in C.lentillifera so far
Additional comments: The first part of the review should be revised in detail, there are some paragraphs where the information should be described clearly. The second part, from "Wastewater treatment by C. lentillifera", requires some minor modification.


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: HIGH TEMPO MUSIC PROLONGS HIGH INTENSITY EXERCISE
Review round: 1
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: Lines 108-109: Please provide references for the termination of the protocol.
Line 187-188: You mentioned gender as a between-subject factor but the potential effects of baseline VO2 consumption was not taken into account. It might be possible that longer exercise duration was because the subjects were more prepared.
Line 145: You mentioned that you altered the tempo to be consistent throughout the playlist. While it might be justifiable to standardize your protocol, it could have affected the experience of listening to music (specially when it includes lyrics). Please explain this further as a limitation.

- Please discuss the clinical significance of measuring ventilation, tidal volume, and RER in your study.
- Please mention whether the number of participants was based on a pilot study (power analysis) and if not, mention it as a limitation and provide suggestions for future studies.