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: PARTICULARITIES OF THE CHANGES IN YOUNG SWIMMERS’ BODY ADAPTATION TO THE STIMULI OF PHYSICAL AND MENTAL STRESS IN SPORTS TRAINING PROCESS
Review round: 1
Reviewer: 3

Output:
Basic reporting: I am only including some basic comments here, as I decided to include the comments directly in the pdf document and upload it due to the large number.
-	From my point of view, important aspects are missing in the introduction, but are addressed in the article in the discussion / conclusion section. In particular, the theoretical background on the interrelationships between recovery and fatigue is missing. Also missing are current results / references to athlete or training monitoring, which can be found, for example, in the consensus paper by Bordoun et al. (2017) or Kellmann et al. (2018).
[Bourdon, P. C., Cardinale, M., Murray, A., Gastin, P., Kellmann, M., Varley, M. C., et al. (2017). Monitoring athlete training loads. Consensus statement. Int. J. Sports Physiol. Perform. 12(Suppl. 2), S2161–S2170. doi: 10.1123/IJSPP.2017-0208
Kellmann, M., Bertollo, M., Bosquet, L., Brink, M. S., Coutts, A., Duffield, R., et al. (2018). Recovery and performance in sport: consensus statement. Int. J. Sports Physiol. Perform. 13, 240–245. doi: 10.1123/ijspp.2017-0759]
-	Some of the statements in the Discussion and Conclusion belong more to the Introduction or Methods chapters.
-	Data should be consistently anonymized throughout (including in the raw data)
-	Abbreviations and designations are partly contradictory or not consistent. For example, four training stages are listed in this study order: E1- basic, E2-specific, E3-pre competitive, E 4- competitive. However, in the associated Table 1, 6 (Prepatory, Specific, pre-Cp., Cp. Transition, Specific) stages are listed for Juniors and 5 (Prepatory, Specific, pre-Cp., Cp., pre-Cp) stages are listed for Cadets.
Experimental design: In my view, the major deficits lie in the description of the study and the methods.
-	It is not explained why two different test procedures are used to measure the same thing (measure the perception of stress: CWT and CPST, competition anxiety: SCAT and CT.  from my point of view, several questions arise here:
1.	what are the differences? How are the tests structured, which items are asked for?
2.	how are the different scores taken into account in the evaluation or the subsequent calculations?
3.	what are the (statistical) quality criteria of the tests? Are validation data available from existing studies?
4.	what exactly is measured with these test procedures? In particular here the theoretical derivation is missing from my point of view.
5.	what is the meaning of the 3 levels or how is the interpretation of the levels, i.e. what effects can be concluded from them on training or performance?
-	How and according to which criteria were the effort zones divided? How was it checked during training that the athletes really swam the specified distance in the respective zone? When and according to which criteria or parameters was it decided whether and how the training had to be adapted? if individual calculations were made for each athlete, why are group values shown in the results? ...
-	Were the anthropometric data measured only at the beginning (as described) or at each stage (E1-E4) (if so, where are the results)?
Validity of the findings: -	Unfortunately, I can't really understand (even from the raw data) how the calculation of the corelations was done. Were the group means correlated in each case or were the individual values calculated and then aggregated?
-	It is also not clear to me on which theoretical basis the assumption is based that mental training indicators (e.g. perceived stress) should have an influence on stature. Likewise, why there should be a correlation between the volume in training zones and the stature?
-	If conclusions are drawn here about the relationship between recovery and fatigue, then this should also be derived in a more differentiated manner in the theoretical justification of the study. In particular, it should be questioned here which definition of recovery and fatigue is assumed. Depending on this, how is recovery or fatigue recorded or measured here in this study? There are numerous articles on this topic, e.g. :
o	Bourdon, P. C., Cardinale, M., Murray, A., Gastin, P., Kellmann, M., Varley, M. C., et al. (2017). Monitoring athlete training loads. Consensus statement. Int. J. Sports Physiol. Perform. 12(Suppl. 2), S2161–S2170. doi: 10.1123/IJSPP.2017-0208
o	Kellmann, M., and Beckmann, J. (eds). (2018). Sport, Recovery, and Performance:Interdisciplinary Insights. New York, NY: Routledge.
o	Kellmann, M., Bertollo, M., Bosquet, L., Brink, M. S., Coutts, A., Duffield, R., et al. (2018). Recovery and performance in sport: consensus statement. Int. J. Sports Physiol. Perform. 13, 240–245. doi: 10.1123/ijspp.2017-0759
-	The aim of the study is to analyze the individual (training) adaptations of the athletes resulting from the different training phases, therefore, from my point of view, it is not comprehensible that the data and calculations are used or presented almost exclusively group based. Therefore the discussion should focus more on individual differences and similarities than on group values or trends
Additional comments: Despite the criticism, first of all, thank you for the elaborate study and quite interesting data and results. From my point of view, however, it is necessary to define the main target more clearly and to prepare the theoretical background accordingly. If the focus is on individual adaptations, which is more than reasonable in the topic complex of recovery and fatigue, the results and analyses should also be individually based. Furthermore, the study design should be described in a consistent manner and all methodological decisions should be described in advance in a theory-based manner (e.g., according to which criteria will be swum in which effort zone). In particular, if interventions are undertaken based on the study results, the criteria according to which decisions are made should be clearly defined.


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: PARTICULARITIES OF THE CHANGES IN YOUNG SWIMMERS’ BODY ADAPTATION TO THE STIMULI OF PHYSICAL AND MENTAL STRESS IN SPORTS TRAINING PROCESS
Review round: 2
Reviewer: 1

Output:
Basic reporting: It is clear the great effort that the authors made in answering all the questions indicated by the reviewers. However, one must consider the authors 'difficulty in transforming the reviewers' indications into material from their own text. For example, there was an indication of several articles, the intention is that these should be incorporated into the introduction or discussion, not just report what has been done. At the same time, it appears that the authors often only copied the reviewers' text, when it was just to exemplify what the reviewers sought to inform or to correct. The article under analysis is complex, at the same time interesting, but the authors must make an effort to leave the text, at the same time, objective, informative and current.
Experimental design: The design is correct, but it is too long. Please describe just the essentials, in this way, the study can be replicate elsewhere.
Validity of the findings: As mentioned before, the sample size is a great limitation considering the validity of the findings. I really think the authors should consider this as a "pilot study".
Additional comments: Please be more consistent in writing, eliminating information that is not really necessary to understand the work.
Very long sentences in English do not work. Try to rewrite in two or three sentences (example: lines 57 to 60).
Along all the text, please revise language and structure.
Line 64: "Otherwise, the stress makes adaptation impossible, the overtraining occurs leading..." It is impossible or hard to make positive adaptations?
Line 89: Is immersion bradycardia necessary for this article?
Lines 111 - 114: It is exactly as one reviewer has written.
Line 128: why is it in the singular first person?
Line 131: Is it Method?
144: Frrestyle is the competitive event, the swimmers are specialists in front crawl stroke.
146: It is body mass (kg), not weight (N).
174: "The individual competitive tests included in the competition calendar " please rewitte, it is not clear.
161: It should be in the Statistical method.
219-222: Why is there an objective here?
Results:
This section, as the study has many variables, is too long. I recommend to reduce the session with less text. Why are there references in this session?
There are six tables in the results, many are difficult to interpret. Consider the possibility of reviewing their format and check if all of them really should be presented or if there is no possibility of presenting more figures as a result. In addition, try to present each Table in a more objective way, without leaving the text repetitive or heavy.
Discussion: I do suggest a short paragraph with the main objetives and summarizing  the main results found instead the first and second paragraph (lines 443 - 460).
Avoid repeat information regarding methods: 461 - 469.
470 - 487: This discussion is intersting, but there is no reference regarding this issue.
Line 496: please review the format of the reference.
I miss a lot of comparisons with other studies, as those indicated in the first review, to clarify the results and to improve the discussion quality.
Conclusion
Please be more objetive.
l


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: PARTICULARITIES OF THE CHANGES IN YOUNG SWIMMERS’ BODY ADAPTATION TO THE STIMULI OF PHYSICAL AND MENTAL STRESS IN SPORTS TRAINING PROCESS
Review round: 2
Reviewer: 2

Output:
Basic reporting: The manuscript presents a meticulous study on the adaptation processes in young swimmers that is very interesting to improve the current academic knowledge regarding the theory and practice of high competition training. I would like to point out that the manuscript has improved substantially and that all the recommendations that were proposed by the reviewers have been correctly addressed and taken into account by the authors. I have reviewed the new final document, and consider that the current quality of the manuscript is sufficient to consider its publication in PeerJ.

I would like to thank the authors for the elegance and good work developed to meet the recommendations of the reviewers. A scientific publication must be the product of the compilation of the work of the researchers and the reviewers of the final document. Science is above all of us and our obligation is to strive to publish quality work. This manuscript comes from this common effort by authors-reviewers, which confers a good document that will awaken the interest of sports sciences. Thanks to the authors for your understanding and consideration of the recommendations.

The manuscript is ready to be published, therefore, I leave the final decision to the editor.
Experimental design: It has been substantially improved.
Validity of the findings: It has been substantially improved.
Additional comments: Dear Authors:

First of all, I want to thank you for your effort and grace in answering the questions raised by the reviewers. The reviewers' job is to collaborate with the authors to improve the final document. Thanks to your excellent consideration this work has been very easy for the reviewers. Second, I want to personally congratulate you on your research work. Sports science must be increasingly based on applicable scientific concepts. And with jobs like yours, we can be closer to this final goal. Kind regards, and let me say that I have very much enjoyed reading your manuscript and the changes you have made to it.

A cordial greeting.


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: PARTICULARITIES OF THE CHANGES IN YOUNG SWIMMERS’ BODY ADAPTATION TO THE STIMULI OF PHYSICAL AND MENTAL STRESS IN SPORTS TRAINING PROCESS
Review round: 3
Reviewer: 1

Output:
Basic reporting: The authors made a great effort to improve the aspects indicated in the previous evaluations. All questions raised have been resolved or answered. The quality of the article has improved significantly.
Experimental design: Properly worded.
Validity of the findings: Limited by sample size, but can be interpreted as initials.
Additional comments: Congratulations on the conduct of the research and on the way of receiving the questions brought by the reviewers.


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: PARTICULARITIES OF THE CHANGES IN YOUNG SWIMMERS’ BODY ADAPTATION TO THE STIMULI OF PHYSICAL AND MENTAL STRESS IN SPORTS TRAINING PROCESS
Review round: 3
Reviewer: 2

Output:
Basic reporting: The efforts to implement the different requirements of the reviewers are clearly visible and well done.
Experimental design: The methods and the study design are transparent and comprehensible as far as possible
Validity of the findings: As already in the first review, from my point of view, an individually based presentation of the data and results would have been desirable. However, I also understand the authors' dilemma that many additional figures would then have been necessary and that different views of the reviewers have to be taken into account.
Perhaps a consideration for the future is to go into more detail on the individual aspects in another article based on selected parameters.
Alternative statistical methods that take greater account of individual differences could then be considered for this purpose (e. g. Bakdash et al. 2017; https://doi.org/10.3389/fpsyg.2017.00456).
Additional comments: - Line 134-137: Delete parenthesis mentioning coaching styles, as it is sufficient to know here that coaching has an impact on athlete motivation. Otherwise, the different coaching styles would also have to be explained.
- Line 136-137: why "artistic swimming sport"
- Line 299-300: name the athletes or abbreviations: "...results, it was observed that 2 athletes (A1, A4) had low values of anxiety, other..."
- Line 312-313 / Figure1: the figure shows the training volume on average per effort zone at the beginning and end of the study. But what do the mean values refer to (training unit, study phase, ...)? This would have to be added to the description.
- Some sentences are too long (e.g. line 418-422), try to shorten or split into two sentences
- Table 6: Please complete the description or legend and match it with the label in the table. For example, only TΨ1 is explained, TΨ2-4 is no longer explained. What do the variables (1-12) in the first column stand for? Overall, the table is very complex and difficult to understand.


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: PARTICULARITIES OF THE CHANGES IN YOUNG SWIMMERS’ BODY ADAPTATION TO THE STIMULI OF PHYSICAL AND MENTAL STRESS IN SPORTS TRAINING PROCESS
Review round: 4
Reviewer: 1

Output:
Basic reporting: see previous review
Experimental design: see previous review
Validity of the findings: see previous review
Additional comments: All questions and comments were clarified or implemented by the authors. Well done.


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: DIGITAL MAPPING OF SOIL TEXTURE IN ECOFOREST POLYGONS IN QUEBEC, CANADA
Review round: 1
Reviewer: 1

Output:
Basic reporting: The paper is clearly written.  I have provided some editorial comments in the general comments section.

The paper is adequately referenced

The figures and tables are fine with the exception of points that I have provided in the general comments section.

No hypotheses were presented in this paper.
Experimental design: The paper describes original primary research and appears to be within the aims and scope of the journal.

The research question in this paper does not seem to be clearly stated.  It is unclear how the research fills a knowledge gap.  The knowledge gap has not been clearly identified.  I think that the authors should be able to more clearly state their research question (s) and identify knowledge gap (s)

The authors appear to have performed a rigorous investigation to a high technical and ethical standard.

The methods are described in sufficient detail. Some comments are provided in the general comments section.
Validity of the findings: The study appears statistically sound.

Lines 404-414. The conclusions from the study seem fairly limited.  This may be related to the relatively limited aim of the study “to map the texture of mineral soil at the scale of the ecoforest polygons”.  The authors should consider ways to make the study appear more than a relatively straightforward mapping exercise.
Additional comments: The authors state on lines 126-127 that “the present analysis aims to map the texture of mineral soil at the scale of the ecoforest polygons in Quebec, Canada.”  The authors used an impressive dataset of 29570 mineral soil samples (17901 sites).  However, the authors describe their study as an exercise in mapping soil texture in Quebec.  It would be good if they provided research questions, objectives or hypotheses and clearly stated the research gap that they are attempting to fill.

Line 187. The authors state that they compiled 33 covariates (and again on line 331), however, the authors should indicate how many covariates they actually used in the modelling.  The number appears to have been much higher due to the creation of dummy variables as described on line 222.  Figure 4 includes “the 50 most important variables”, suggesting there were many more than 33 covariates used.

Lines 190-192.  The authors need to clarify in detail what they mean by “estimable”.  They need to state exactly how they obtained the covariate data.

Line 201 It is not clear what the authors mean by geological material.  This is not the correct terminology.

Lines 186-211.  The authors need to clearly state the sources of their covariate data.  For example, where exactly is the “soil physical environment” data mentioned in line 202 from. This needs to be clarified.  Also, the term “soil physical environment” is potentially misleading and should be changed to something such as “soil texture and drainage class”.
The authors refer to “soil physical environment” in line 202-203 and in Table 1 they describe the covariates used as “Soil physical environment: very shallow (< 25 cm) or very stony (0); coarse texture and xeric to mesic drainage (1); medium texture and xeric to mesic drainage (2); fine texture and xeric to mesic drainage (3); coarse texture and hygric drainage (4); medium texture and hygric drainage (5); fine texture and hygric drainage (6); hydric drainage (7); organic (8: Fen, 9: bog).”  Was this information collected in the field or obtained from another source?  This needs to be clarified.  It appears that possibly, the information was included in the ecoforest polygon mapping, but this needs to be clarified.

Lines 358 and on.   It appears that the authors used mapped texture class (coarse, medium, fine) information as a covariate in their modelling.  They then found that the most important variables in their models were variables “Describing soil physical environment and soil surface deposits” (line 359).  In particular, SoilPhysEnv 1 and 6 as indicated in Figure 4 were found to be important and these are” coarse texture and xeric to mesic drainage (1); fine texture and hygric drainage (6)” as indicated in Table 1.  This seems to suggest that the authors used estimates of soil textural class from the ecoforest polygon mapping as a covariate for their modelling.  Essentially, it seems that maps of soil texture in categories of coarse, medium and fine texture could be produced directly from the ecoforest polygon maps prior to this study, and that what the authors have done is refined this mapping to produce maps of soil texture with more detail.  This should be clearly stated and discussed in the paper (introduction, methods, results, discussion, conclusion).

Line 373-374 A reference is needed here.
Figure 1. there should be some labels on the map to orient the reader, such as the border of Quebec, location of a few major cities, and names of water bodies

Minor Editorial Comments
Line 25 “on the field” should be “in the field”
Line 78.  The point is not stated as a technical limitation.
Line 155  “sylvicultural” should be “silvicultural”
Line 165 “”the mixed forest” should be “in the mixed forest” and “the continuous boreal” should be “in the continuous boreal”
Line 197 “tiles” should be “tile”
Line 214 “totalize” is not a word
Line 230 “others” should be “other”
Line 354 “from entire region” should be “from an entire region”
Line 377 What is meant by “experimented photo-interpreters”?
Line 387 “a large soil sample datasets” should be “large soil sample datasets”
Line 413-414  The meaning of the last sentence in the conclusion is unclear.


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: DIGITAL MAPPING OF SOIL TEXTURE IN ECOFOREST POLYGONS IN QUEBEC, CANADA
Review round: 1
Reviewer: 2

Output:
Basic reporting: See General comments for the author below.
Experimental design: See General comments for the author below.
Validity of the findings: See General comments for the author below.
Additional comments: This is a well-written and well-structured manuscript that applies machine learning to map soil texture in a large ecoforest region in Quebec, Canada. The approach is fairly standard but it is state-of-the-art and has some novel elements too. The methodology is overall sound but not always clearly explained and may be criticised on some aspects. My concerns are worked out in the detailed comments below.

Detailed comments

(L97-107) It is not about whether a vector or raster representation of covariates is used, these are just technical aspects that do not truly influence the statistical modelling. In the end it is about how much of the soil variation is explained by the covariates, regardless of whether they are polygon or raster maps. What is important is whether soil variation is considered abrupt or gradual. Authors assume that soil spatial variation within polygons is small compared to variation between polygons, but is this true? This is just an assumption as stated in L102, so I don’t think you can maintain the general claim in L105-107.

(L108-119) Clearly a lot of effort has gone over the past 50 years into producing the ecoforest polygon map and indeed it turns out to be a valuable source of information in this study. That in itself is very fine but note that it is not a solution in cases where such information is not available. In other words, the approach used in this work cannot easily be extrapolated to other studies. Perhaps this could be mentioned in the Discussion. Authors might even check whether detailed polygon maps are available for other parts of the world and in this way discuss the extrapolation potential of their method. It was also not clear to me how the polygon maps were used in this study. Were only the polygon delineations used, by averaging covariates within polygons (L191-192 seems to suggest this, it is also apparent from Figure 7 that shows abrupt changes in texture predictions at polygon boundaries)? Or were the attribute values of the polygons (L111-113) used as covariates? I did not see these mentioned clearly in Table 1, so perhaps they weren’t? If not, why not? If they were used, there may be a problem that the soil observations used for calibration and cross-validation were also used to create covariates, which leads to overoptimistic validation metrics.

(L135-136) I did not come across the term ‘soil province’ before, is this a common term? If not, please define it.

(L143) There are more soil samples than sites, is that because sometimes there more than one observation per profile or is that because there are multiple locations per site. I think it is the former, because later on authors explain that samples from the B and C horizon were taken, but here I was not sure. Perhaps it is because of the term ‘site’ which is larger than a point location. For the PSPs the sites are 400 m2 circular plots (L146), so how was the soil sampling done? Were these soil samples from the centre of the circle or composite samples from multiple points within the plot? This should be clarified, also for the other two datasets (EOPs and SIP). Since soil data from three different programs were merged, it is important to describe any possible systematic differences between field and lab work between the programs, and whether this may have affected the results. L181-185 indicates that all soil samples were treated exactly the same way, over a very long period of time. Is this true?

(L156, L169-170, L178) Soil samples were taken from the first diagnostic B and C horizon. These were all merged into a single dataset but in the modelling the horizon was included as a covariate (Figure 4, Table 1). So when the calibrated model is used for prediction we may predict texture both for the B and C horizon. Figures 6 and 8 refer to the B horizon (Figure 7 presumably too, but this is not specified in the caption). What about the texture maps of the C horizon? And how to interpret the B horizon texture map for locations that have no B horizon (this happens, see L178-179)? Note that fitting a single model that assumes that a single binary covariate “B or C horizon” explains the difference between texture in the B and C horizon is quite rigid, what about fitting separate models for the B and C horizons? Was this tried and if yes how well did it predict soil texture? I would also like to see summary statistics of the top, bottom and thickness for both horizons, how variable are they? In the end we get maps that predict soil texture for horizons, but for users such as provincial forest managers, such maps may only be useful if it is also known what the depth intervals of the B and C horizon are at every prediction location. Are the B and C horizon indeed most interesting for forest managers, more so than the A horizon? Can they reliably be sample using an auger (L156)? Why was it done this way instead of mapping soil texture at fixed depth intervals (this is what most studies do, including the Hengl. et al (2017) study that authors use for comparison)? Soil samples were taken in one-third of the EOPs, was this subset chosen randomly from all EOPs?

(L171) Only 9476 horizons of 7085 sites were sampled. Why not 2 x 7085? Is this because for many sites there was no diagnostic B horizon? But this would mean that diagnostic B horizons are quite rare? Is this not a problem?

(L178-180) The text suggests that for each site no more than one soil sample was taken, but we still have 839>806.

(L201-203) This suggests that geological covariates were measured at the sampling sites, but these are not known at prediction locations (unless they are interpolated in some way, but this introduces uncertainty) so cannot be used to calibrate the machine learning model. Please clarify. It may be that all covariates were averaged over the polygons before they were offered to the machine learning model, but this was not clear to me. If this was done, then state this more clearly. Also, why would this be a proper way of preparing covariates? It is built on the assumption that soil and covariate variation within polygons is negligibly small, but is that really the case? Why would it be needed, why not use the covariates as raster maps? Maybe I simply did not get it, but in a way this then indicates that authors can do a better job in explaining how the polygon approach was applied and why it is useful.

(L205-209) Why refer to ecological regions based on physical environment, climate regimes, soil provinces and vegetation as ‘spatial position and context’? Are these not covariates that refer to environmental conditions (i.e., climate, vegetation, terrain)?

(L228-231) The type of machine learning algorithm and the machine learning hyperparameters were also optimised. What dataset was used for that? To guarantee independence, the data used for this should not also be used for model performance evaluation. In other words, a nested cross-validation approach should be used, where data are split in three parts (calibration, validation and test data).

(L234) Does the R-square refer to that computed on the 1:1 line or on a linear regression of predicted on observed (i.e., the squared Pearson correlation)? This should be clarified, and in fact the first of these should be used. To avoid confusion it would be better to refer to it as ‘model efficiency coefficient’ (for example see https://en.wikipedia.org/wiki/Nash%E2%80%93Sutcliffe_model_efficiency_coefficient).

(L243, L321-328, L389-402) Authors quantified prediction uncertainty of V1 and V2 but what we really want is the uncertainty of the clay, silt and sand predictions. Perhaps they could try and compute these. This requires that the correlation between the V1 and V2 prediction uncertainty is quantified (can be done, see https://arxiv.org/abs/2005.14458) and next that an uncertainty propagation analysis is done (Monte Carlo simulation is easy but computationally demanding, perhaps it can be done analytically or using a Taylor series approximation approach). Perhaps it is too challenging, but if they succeed it would be a very worthwhile extension of the methodology. If not feasible, it may be mentioned as a useful topic for future research in the Discussion. As it is now, Figure 8 and L321-328 present and discuss the V1 and V2 uncertainty maps but can these be easily interpreted in terms of texture, given that the transformation from V1 and V2 to clay, silt and sand is highly non-linear?

(L245-251) Not very clear what was done here, it suggests that in the end predictions were made on a grid after all. So where does the polygon approach go? Was that only used to average covariates within polygons (which may be criticised, see comment above)? And what are ‘regional polygon maps’? How do these differ from the ecoforest polygon map introduced in L108?

(L279-280) That depends on the nugget-to-sill ratio: if this is small, I would not conclude that there is a high amount of short-range variability. Please reformulate.

(L287-319) I like that authors pay ample attention to interpretation of the resulting maps and explain the patterns that were obtained from a pedological perspective.

(L333-341) Interpretation and comparison of the proportion of variance explained is tricky for study areas that are not the same because it is a ratio and depends on the amount of variance present in the study area. For example, for global mapping as done in Hengl et al. (2017) it is relatively easy to get a high R-square because the denominator of the ratio may be larger for global data than for local/regional data. A more honest comparison would be using the RMSE or MAE.

(L350-356) The abrupt spatial transitions in Figures 6 and 7 is simply the result of the modelling approach used here (if I am right authors averaged covariates within polygons before using them to train the machine learning model). The text suggests that this is a good thing but it is merely a result from the choices made, and it was not shown that his is better than alternative approaches that do not pre-process covariates. Authors should prove that their approach is better by comparing it with a method that trains a machine learning model on the same covariates, but then without first averaging within polygons, using the same training data.

(L383) It seems to me that the computing costs of residual kriging would not be that demanding in this study, if at least authors would limit the conditioning data to data within a local neighbourhood. Why not do this and check out the improvement?

(L410-411) That really also depends on the accuracy of the map. Different uses require different accuracy levels, so substantiate this claim by quantifying the uncertainty of the  prediction maps and evaluating if these are within limits required by provincial forest managers.

(Figure 2). Explain the horizontal bands in these figures. The straight blue line supposedly represents the linear regression but it is very close to the 1:1 line. I find this hard to believe. It should have a much steeper slope (because when models explain only part of the spatial variation, predictions are smoother (have lower variance) than observations. Same problem occurs (but to a lesser extent) in Figure 3.

(Figure 5) Is it realistic to assume that residuals in the B and C horizons have the same variogram? This goes back to the fairly rigid assumption that largely the same model may be used for both horizons (same holds for Figure 4, also here the importance of covariates is forced to be te same for the B and C horizons). Perhaps more accurate models can be obtained if a more flexible model was used (separate models for B and C horizons).

(Figure 7) It may be useful to indicate this area by a rectangle in Figure 6. What do scale 1:200,000 and scale 1:100,000 refer to? If I am right the lower panel is no more than another zoom in on a rectangular area in the top panel. Better avoid the scale terminology. From the bottom panel it also seems that there is some spatial variation in predictions within polygons, although I am  not certain because I do not know where polygon boundaries are. If this  is true, what is the cause of this (assuming covariates were constant within polygons)?


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: DIGITAL MAPPING OF SOIL TEXTURE IN ECOFOREST POLYGONS IN QUEBEC, CANADA
Review round: 1
Reviewer: 3

Output:
Basic reporting: Digital mapping of soil texture in ecoforest polygons in Quebec, Canada

Dear Editor
I have read above article very carefully. I have the following major comments.

Abstract
Which models?
Please add variables in this part.
Any keywords?
Introduction
Please order citations from old to new.
Literature review needs to some new citations during 2019-2021.
Introduction is written too long.
Please explain about innovative of this study.

Study area
Please add coordinate system of the study area.
Material and methods
Please add a flowchart for the current study.
Covariates
It is important to explain about the applied variables and explain about preparing these factors.
What is its scale or spatial resolution?
Any multi-collinearity test on the applied variables?
Results
This part is written so poor and weak.
Discussion
It is important to add some comparisons with previous publications, advantages, disadvantages, suggestions and future tasks.
Experimental design: please consider my comments.
Validity of the findings: please consider my comments.
Additional comments: Digital mapping of soil texture in ecoforest polygons in Quebec, Canada

Dear Editor
I have read above article very carefully. I have the following major comments.

Abstract
Which models?
Please add variables in this part.
Any keywords?
Introduction
Please order citations from old to new.
Literature review needs to some new citations during 2019-2021.
Introduction is written too long.
Please explain about innovative of this study.

Study area
Please add coordinate system of the study area.
Material and methods
Please add a flowchart for the current study.
Covariates
It is important to explain about the applied variables and explain about preparing these factors.
What is its scale or spatial resolution?
Any multi-collinearity test on the applied variables?
Results
This part is written so poor and weak.
Discussion
It is important to add some comparisons with previous publications, advantages, disadvantages, suggestions and future tasks.


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: DIGITAL MAPPING OF SOIL TEXTURE IN ECOFOREST POLYGONS IN QUEBEC, CANADA
Review round: 2
Reviewer: 1

Output:
Basic reporting: see general comments
Experimental design: see general comments
Validity of the findings: see general comments
Additional comments: The authors have addressed the concerns of the reviewers adequately with one exception which follows:

The authors don’t appear to have carried out any form of external validation of their model.  They carried out internal validation using 5 repeats of 5-fold cross-validation, but this was used to tune parameter values.  They should  have used a test data set that was not used in the internal validation.  Ideally, the authors should validate their model with an independent dataset, however, since this is likely not feasible at this point, they should discuss this limitation of their study in their discussion section.


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: DIGITAL MAPPING OF SOIL TEXTURE IN ECOFOREST POLYGONS IN QUEBEC, CANADA
Review round: 2
Reviewer: 2

Output:
Basic reporting: See below.
Experimental design: See below.
Validity of the findings: See below.
Additional comments: I am satisfied with most of the authors’ responses to my comments to the original manuscript and how they incorporated these in the revision. I do not agree with everything they write, but in many cases this is a matter of opinion and I accept that different views exist. I did get the impression that authors are sometimes too easily satisfied and could have done more with my suggestions.

There are some points that I do need to mention explicitly and that in my view may call for a new revision round:

1.	I still have serious concerns about how authors dealt with the vertical dimension (depth) in this study. For example, Figure 7 shows maps of the soil texture composition of the diagnostic B horizon but large parts of the study area do not have a diagnostic B horizon. So how to interpret this map? How can one predict soil texture of a horizon that is not there? The escape route that authors use in their response is that in fact soil texture does not vary much with depth so it really does not matter that much at what depth soil texture is predicted, and whether it is the B or C horizon or any other horizon. I find this unsatisfactory, much too easy, and is it really true? If that were the case why not forget about depth altogether, ignore horizons and simply predict in 2D and use calibration data from all depths? Can authors provide some evidence that shows that vertical variation in soil texture is negligible (note that they write themselves that clay can accumulate)? Please note that I am not asking that authors do a 3D modelling exercise because I fully agree that having observations at only two depths per sampling location is not enough for calibration of a 3D model. But there are some important methodological problems with predicting soil texture for the diagnostic B and C horizons, as done in this paper.

2.	“For some unknown reason, 33 sites have 2 samples in the database” (L194). Is this not very worrying? Should you not check why this is the case? After all, you selected observations from the first diagnostic B soil horizon only, so there can be only one? Or were duplicates taken, to quantify micro-scale variation and/or lab errors? I would want to know what is the cause of 33 sites having 2 samples in the database because it may also point to serious errors in the database (wrong attribution of soil samples to sampling locations). Why are authors so easily satisfied, why don’t they go to the bottom of this?

3.	In their response authors make clear that the soil characteristic covariates were measured at sample sites and characterized by photointerpreters at the prediction locations. This is worrying too, because it effectively means that different covariates were used for calibration than for prediction. For prediction, we have only expert judgements of the ‘true’ covariates, and experts make errors (and must ignore spatial variation within polygons). All this affects the results, in particular the uncertainties (prediction intervals). These will be too ‘optimistic’ because the model assumes that the ‘true’ covariates are known at prediction sites while all that we have are proxies of them. For a paper that addressed this problem in a case where the covariate was soil type (as measured on sampling sites, as depicted on a soil type map at prediction sites), see model 3 in Kempen et al. (2010), European Journal of Soil Science 61, 333-347.

4.	Authors confirm that the R-square is defined as the square of the Pearson correlation. I find this very unfortunate, because this does not convey how close the predictions are to the observations. For instance, in all cases where prediction=a+b*observation we will get an R-square of 1, regardless the value of a and b (except when b=0). So even if there is a strong bias (intercept a different from zero) and smoothing (slope b smaller than 1) we still would conclude that the model performance is perfect. Clearly undesirable. Authors defend their choice because this is how it is done in the postResample function of the caret R package, but do they seriously believe that this is a valid justification? If others do things wrongly then they can do it as well? Again I think authors are much too easily satisfied. Instead they should help correcting this mistake and clarify to the DSM research community that in case of (cross-)validation, where we compare predictions with observations, performance should be evaluated against the 1:1 line instead of against the fitted linear regression between predicted and observed. For this we can use Lin’s concordance correlation or the Model Efficiency Coefficient, as I had suggested in my review of the original manuscript. Perhaps in this study the differences may not be very large, but in other cases it will and we should not misguide the readers.

5.	Authors prefer to keep the reference to cartographic scale in Figure 8 because “this is a common element of a map”. They explain it now and write in the caption that scale 1:200,000 means that 1cm on the map represents 2km on the ground. So when the article is published, should I measure the distance in cm on the computer screen to derive the distance on the ground? What if I zoom in on the PDF? What if I use a different computer with a bigger screen? I am sorry for being cynical, but if authors had given this a bit more thought they would have realised that cartographic scale only works well with printed maps.


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: DIGITAL MAPPING OF SOIL TEXTURE IN ECOFOREST POLYGONS IN QUEBEC, CANADA
Review round: 2
Reviewer: 3

Output:
Basic reporting: Dear Authors
Thank you so much for revised version. I have read considered the revised version. I am positive with this version. My decision is accept in the current form.
Experimental design: Experimental desinged very-well.
Validity of the findings: good.
Additional comments: Thanks for revised version.


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

Input:
Title: DIGITAL MAPPING OF SOIL TEXTURE IN ECOFOREST POLYGONS IN QUEBEC, CANADA
Review round: 3
Reviewer: 1

Output:
Basic reporting: Fine
Experimental design: Fine
Validity of the findings: Fine
Additional comments: I suggest the paper now be accepted.


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: DIGITAL MAPPING OF SOIL TEXTURE IN ECOFOREST POLYGONS IN QUEBEC, CANADA
Review round: 3
Reviewer: 2

Output:
Basic reporting: See below.
Experimental design: See below.
Validity of the findings: See below.
Additional comments: I am happy with this second revision and response to my comments and have no further 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: THERAPEUTIC EFFECT OF AND MECHANISMS UNDERLYING THE EFFECT OF MIR-195-5P ON SUBARACHNOID HEMORRHAGE-INDUCED VASOSPASM AND BRAIN INJURY IN RATS
Review round: 1
Reviewer: 1

Output:
Basic reporting: The manuscript should be edited by a native speaker of English. Additionally, the raw data of immunobolt was missing.
Experimental design: Methods failed to offer sufficient information to be reproducible by another investigator.
Validity of the findings: The raw data of immunobolt was missing.
Additional comments: In the current study, Tsai et al investigated the role of miR-195-5p in the pathological process following subarachnoid hemorrhage (SAH). And they concluded that administration of miR-195-5p alleviated SAH-induced vasospasm and brain injury. It’s an interesting study. However, several issues must be addressed.
1.	Method. How did the author choose the delivey route and dose of miR-195-5p in this study?
2.	Method. Had the author determined the concentration of miR-195-5p in brain?
3.	Method. Did the authors perfuse rats with formalin before the basilar artery morphometric analysis?
4.	Method. How many animals were used for each test, including Basilar Artery Morphometric Analysis, Cell Death Assay, immunoblot, and TNF-α evaluation? They wrote that 96 animals for each test, which was quite complexing.
5.	Method. As mentioned in Statistical Analysis, the difference among the groups were analyzed by ANOVA. How were the data analyzed to ensure they were normally distributed, a requirement of ANOVA?
6.	Data points should be indicated on all statistical graphs. Providing the bar graph and only statistical parameters can suggest that the data underlying any particular bar are normally distributed and contains no outliers, when this may not be the case.
7.	Generally, microRNA selience specific gene through binding to the DNA fragments. How could the miR-195-5p modulate the level of eNOS and iNOS, as well as regulate the phosphorylation p-65 and IκBα?


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: THERAPEUTIC EFFECT OF AND MECHANISMS UNDERLYING THE EFFECT OF MIR-195-5P ON SUBARACHNOID HEMORRHAGE-INDUCED VASOSPASM AND BRAIN INJURY IN RATS
Review round: 1
Reviewer: 2

Output:
Basic reporting: This study aimed to determine the effects of miR-195-5p on an experimental model of SAH in male rats. It was unclear why authors selected this specific miRNA for studies. No clear rationales or justifications were provided except listing a couple of studies showing beneficial effects of miR-195 mainly in cancers. No prior studies that associate SAH with miR-195 were provided to support the scientific premise of the study.
Experimental design: The miR-195 and control were in suspensions delivered in tail vein injections. It is questionable how this method could ensure BBB penetrability and exhibit efficacy on brain changes as authors assumed. The small RNAs are unstable and peripheral delivery requires carriers to prevent degradation before getting access into the brain. No validation studies were provided about whether miR195 was efficiently getting into the brain and increasing the levels in the brain. With this questionable method for studies, the validity of findings is questionable.
Validity of the findings: see above. The validity of the findings is questionable.
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: SPATIAL-TEMPORAL VARIABILITY OF MYTILUS GALLOPROVINCIALIS LAMARCK 1819 POPULATIONS AND THEIR ACCUMULATED SEDIMENT IN NORTHERN PORTUGAL
Review round: 1
Reviewer: 1

Output:
Basic reporting: The English needs major revisions by a mother tongue. I made some comments below but there are many more examples and some sentences are hard to follow, making the manuscript sound in part repetitive and redundant, and in part confusing the reader that has to go over the same sentences many times before grasping the meaning.

The introduction is generally too long and while it provides a wide range of references (although none very new) and ideas , it seems largely unfocused jumping from the importance to some various threats and only living a little space to the study itself. I wonder if it could be more focused to the key questions and gaps.
The discussion can also be more structured
The conclusion is also unfocused, with repetitions of results that should not be in a conclusion section.
Experimental design: This is a well designed monitoring study on population dynamics of Mytilus galloprovincialis in Portugal.
Unfortunately it lacks essential complementary environmental metrics - especially with regards to hydrodynamics, which makes the study weaker as not much can be explained and thus the discussion is full of speculative sentences.
It is a shame because it is very interesting that there are not many seasonal effects, especially with regards to spat or loss of cover from wave action (although i do not understand whether the shores are very exposed or locally sheltered).
Validity of the findings: The study could be better introduced and discussed in a wider context. It is lacking of a strong focal discussion point.
More environmental data to relate these findings could be useful and an additional assets.
In alternative I suggest to revisit some aspects related to explaining the findings, since there are no data: for example the byssal strength in relation to waves cannot be explained simply by the thickness of the clumps. So is the optimal temperature theory that is not tested and it comes out as a strong result according to the conclusions.
Additional comments: abstract
L22 Marine repeated two times
L27 descriptors is vague here
L 28 what does along the year mean? Better to esplicitate the cadence of sampling
L 30 results showed
L33 lack of seasonality for what?

introduction
L 44 I would refrain from using subjective words such as ‘important’
L 45 they can control - it is too ‘active’ and they don’t control it on purpose
L 53 probably as a result?
l 56 I am not sure that they have such a high value. they are much less costly than other bivalves-
L 56 it is vulnerable to rather than exposed
L 59 see major comment about English

L 78 what is it meant by high water circulation? Strong flow rates / wave action?
L 79-83 English

methods
l170 if i understand correctly winter was split in two ‘winters’ (beginning of the year and end of the year) - this should be better rationalised in the methods

discussion
here also the english should be revised throughout
the reference in L 324 should read Ceccherelli

L 360 - were the clumps studied in exposed parts of the shore? and byssal strength was not measured. It is not necessary that thickness increases but they may produce more threads

L 387 - can none of this be tested?

L 408 is there no model of wave action locally or the possibility of adding dynamometers on the shore?

Figures: are the letters to indicate significant differences between sites also there to indicare significant differences between months ?

Raw data - JaAg12: sample in January, in Aguda, shore 1, site 1 I think this should be site 2? If understand from the other examples


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: SPATIAL-TEMPORAL VARIABILITY OF MYTILUS GALLOPROVINCIALIS LAMARCK 1819 POPULATIONS AND THEIR ACCUMULATED SEDIMENT IN NORTHERN PORTUGAL
Review round: 1
Reviewer: 2

Output:
Basic reporting: The Results section is quite lengthy for reading. Repetitive by saying that "this statistically significant", when a more integrative description if preferred. Small statistical differences are not biologically relevant, and authors should only emphasize those of interest for their work.

The Discussion section is very long, considering obtained results. Authors should try to integrate the results obtained for each endopoint and not describing them individually.
Experimental design: The authors separate samples in two season blocks: autumn-winter and spring-summer. This makes that January 2019 is grouped with October and December 2019. That is, months separarted almost for one year are grouped. Authors do not provide information regarding environmental variables at each month, that allow to group them together. Only a mention to two previous works is done in Lines 150-153. Environmental data collected at each month should be also included as Supplementary Table or in the Raw data file.

In the methods section a Table should be included describing the characteristics of each sampling site. Sediment type (rocky, sandy, muddy..), existing impacts (industries, urban, agricultural, protected área..). This allows a better knowledge of the environmental conditions existing at each site.
Validity of the findings: The absence of significant seasonal or site differences regarding biological endpoints is very odd. Authors mention that previous works in the same area described seasonal changes in mussel physiology. Considering that mussels in the North West Iberian Peninsula spawn during spring and summer time, it is not clear why the mussels analyzed in the present study showed a constant condition index.

It is well known that mussels inhabiting environments rich in nutrients can produce gametes all along the year. Nutrient sources could be natural (constant phytoplankton production) and/or artificial (organic materials arriving from rivers or urban pollution). Author mention that changes in phytoplankton content were previously described in the study area (Lines 159-160 and 379-383), but overall they assume that nutrient status is optimal all along the year in their mussel populations. What about organic material pollution sources? Could they contribute to the constant condition indesx determined in studied mussel populations? are the CI values determined in those mussels high, medium or low in comparison to data existing in the literature?
Additional comments: The manuscript is well written and clear. The organization of the contents is also correct. The experimental approach is correctly described. A more detailed information regarding sampling points should be included in the Study Area section.
Some of the results, biological endpoints mainly, need some more details. For example the absence of changes in CI requires a better interpretation and find an association with existing environmental variables.


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: SPATIAL-TEMPORAL VARIABILITY OF MYTILUS GALLOPROVINCIALIS LAMARCK 1819 POPULATIONS AND THEIR ACCUMULATED SEDIMENT IN NORTHERN PORTUGAL
Review round: 3
Reviewer: 1

Output:
Basic reporting: The article has improved, it is now easier to read and follow through. I appreciate the authors effort.
Experimental design: The authors have made the corrections suggested, including adding environmental data.
Validity of the findings: The addition of the conclusion is appreciated, despite this is 'speculative' it is interesting for future research.
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: SPATIAL-TEMPORAL VARIABILITY OF MYTILUS GALLOPROVINCIALIS LAMARCK 1819 POPULATIONS AND THEIR ACCUMULATED SEDIMENT IN NORTHERN PORTUGAL
Review round: 3
Reviewer: 2

Output:
Basic reporting: This interesting paper tries to fill knowledge gaps of spatial-temporal variability of Mytilus galloprovincialis populations along the Atlantic coast of the Iberian Peninsula and the accumulated sediments on their clumps.

The title clearly reflects the contents and the abstract is sufficiently informative.

I am not a native English-speaking person, but I think the language and grammar are clear.

The literature cited is well referenced and relevant.

Introduction
I have included some minor corrections in the attached PDF.

Lines 65-71 - I suggest expanding this paragraph where the effect of environmental factors on mussel populations is mentioned since this is the main objective of the paper. Also, provide more information about the interest of evaluating the accumulation of sediments on these coasts, that is, if it is because a natural accretion-erosion process affects them or if it is due to increases in turbidity due to anthropic activities

Figures
Figure 1- I suggest improving the quality of this figure and include the map coordinates.
Figure 2 – I suggest change the label for this sentence: Annual variability of environmental variables registered along the year at the studied area. (A) Temperature during 2019. Red and green lines show the range of tolerance and optimal temperature of Mytilus galloprovincialis respectively. (B) Salinity during the year 2018. (C) Wave height during 2019.
Figure 3- Missing space between words Mean// values and  of//percentage
Figure 4, 5, 6 and 7 – I suggest bring the y axes of all the figures to the same maximum value.
Figure 8 - Missing space between words Mean// values. Check the title of the y-axis of figure 8A.
Experimental design: In the description of the study areas, it is necessary to clarify why these beaches were selected as different factors for the analysis. It is if the chosen beaches are under different anthropic pressures (harvesting, trampling, etc.), or have some particular environmental characteristic. Sampling and Data analysis are clear and sufficient to support the research.
Validity of the findings: Discussion

Line 280- Please expand this sentence, indicate if there are differences in anthropic pressure between beaches. What indicators allow you to reach this type of conclusion?

Lines 298-300 -I do not agree with this explanation. For this study, wave measurements were not taken at each beach, there is only a general wave record from a buoy. Although this wave record indicates that wave height varies seasonally, the intensity of the waves at each beach is likely to be different due to local shoreline characteristics.
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: HOST MIGRATION AND ENVIRONMENTAL TEMPERATURE INFLUENCE AVIAN HAEMOSPORIDIANS PREVALENCE: A MOLECULAR SURVEY IN A BRAZILIAN ATLANTIC RAINFOREST
Review round: 1
Reviewer: 1

Output:
Basic reporting: Brazil has tremendous biodiversity, and many understudied ecosystems. With over 1800 bird species, it is one of the nations richest in avifauna. Within these birds are their blood parasites, of which, with molecular methods, the scientific community is discovering a large number of undescribed species. At this point, in the MalAvi database, hundreds of haemosporidian cytochrome b lineages have been described from Brazil. Many of these were identified by the group of Erika Braga, who presides over the work presented here in the manuscript “Host migration and temperature influence avian haemosporidians prevalence: a molecular survey in a Brazilian Atlantic Rainforest”.  The work describes the parasites of over 1400 birds tested from the Restinga habitats of Northeast Brazil. These coastal forests are threatened by human development, and they harbor some endemic bird species. Of the 70 species of birds studied here, the authors found the overall prevalence of Haemoproteus and Plasmodium to be 22.3%. Interestingly, they found that compared to resident birds, migratory birds have a lower prevalence of Haemoproteus. They also found an association of Plasmodium prevalence with temperature.  However, other traits, such as age, presence of ectoparasites and breeding condition were not associated with parasite prevalence. They identified 8 new cyt b parasite lineages.  The large-scale sampling regime is particularly impressive. The work is relatively straightforward and will be recognized as providing baseline prevalence data for birds of this region of Brazil. I have a few comments that would strengthen this manuscript.
Experimental design: 1.	It would be important to mention why blood smears were not made in this study. Providing data from blood smears would help with the description of new species, and also contribute information about parasitemia. Perhaps due to limited resources, it was not possible to make blood smears, but this should be discussed.

2.	The beginning of the Results section is somewhat confusing. The authors found 322 individuals infected with Plasmodium or Haemoproteus but in the next sentence “All samples that screened positive were subjected to the cytochrome b PCR, which successfully amplified infections from 145 individuals”.  It would be important to clarify why the two different PCR methods should yield such different results. Perhaps in the Discussion, you could point out why you chose to use two different PCR assays, and the advantages of each.  Can you actually trust that the first 322 were actually positive?  Please provide information about how many times the initial PCR was done, just once, or repeatedly. This type of information is directly pertinent to the validity of prevalence data. Also, in line 274, 15 of the 322 infected host individuals (4.6%) exhibited multiple infections. In fact it would be more correct to state that 15 of the 102 sequenced individuals were multiple infections, since the prevalence varies so widely with the PCR methods. Another reason to collect blood smears is to verify PCR results on prevalence.

3.	In line 303, it is not clear why the number of birds decreased from 1443 to 1187 for the final dataset. Was it because these were recaptures, or was there not enough blood? Please clarify.

4.	Temperature can vary widely at the microclimate level. I suspect that the INMET temperature data would be very broad. In the future, in taking temperature measurements, it would be more helpful to take more localized temperature readings. In any case, it is difficult to evaluate how temperature affects the prevalence, because in most cases, the haemosporidian infections are chronic, meaning that the bird will harbor the parasite for many months, or years. It is difficult to determine in what season the birds were infected. For these reasons, it would be helpful in the Discussion to go into more depth relating the data here to previous studies where temperature correlated to Plasmodium prevalence. You already include the references by Ishtiaq, 2017 and LaPointe, 2010. There are some more that are pertinent to malaria transmission in general. For example:
a.	Sehgal, R. N. M., et al. "Spatially explicit predictions of blood parasites in a widely distributed African rainforest bird." Proceedings of the Royal Society B: Biological Sciences 278.1708 (2011): 1025-1033.
b.	Mordecai, Erin A., et al. "Thermal biology of mosquito‐borne disease." Ecology letters 22.10 (2019): 1690-1708.
c.	Fecchio, Alan, et al. "Climate variation influences host specificity in avian malaria parasites." Ecology letters 22.3 (2019): 547-557.

5.	Similarly, it would be helpful to have more lengthy discussion about how the results with migration, age, breeding condition and ectoparasites corroborate or differ from previous studies.

6.	Lines 283-287: These relationships between Plasmodium and Haemoproteus have been established many times. This can be omitted.

7.	In general, the English writing could be improved. I will leave this to the editors, but a couple spelling mistakes: Line 391- whole (not hole).  Line 81-spatial (not spacial).

8.	Figure 1: Please explain in the Figure Legend the different colors of green shading in the “Sampling Area”.

9.	Figure 2: It would be very helpful to include well-characterized morphospecies in this phylogenetic tree. For example, P. relictum, P. circumflexum…. and also several of the well described Haemoproteus lineages. This would help put this tree into context with other known studies about phylogenetic inferences of these parasites.

10.	Typically these types of studies also include Leucocytozoon lineage data. You could briefly discuss why those data were not included in this work.
Validity of the findings: no comment
Additional comments: In sum, this study adds to the literature about the overall lineage diversity of haemosporidian parasites. The sampling was thorough, and the number of birds tested is impressive. It would be helpful to put the work more into context by describing the special characteristics of the Restinga habitats, and the rarity of the samples. Although the use of the different PCR methods is somewhat confusing, and a lack of blood smears problematic, the work provides important information about the diversity of these parasites in a threatened region of Brazil.


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: HOST MIGRATION AND ENVIRONMENTAL TEMPERATURE INFLUENCE AVIAN HAEMOSPORIDIANS PREVALENCE: A MOLECULAR SURVEY IN A BRAZILIAN ATLANTIC RAINFOREST
Review round: 1
Reviewer: 2

Output:
Basic reporting: English is in general good, but several parts need rewriting.

A few extra references could be added in 2-3 specific places mentioned below.

Professional article structure.

Self-contained and relevant results.
Experimental design: Everything appropriate except that some parts of the methods need to be improved.
Validity of the findings: The finding that migration is an important factor is based on an odds-ratio of 0.03, which leaves space for criticism.

I required an improvement of the final sentence of the Discussion and a broader approach than only Brazil.
Additional comments: The manuscript provides information about avian haemosporidians from a novel region the restinga of northeast Brazil. Total sample size is good collected from dozens of species during a two-year period. The manuscript is in general well written, has an almost adequate cover of the literature and provides new insights about the theme. Several parts of the Methods need clarification. Though I liked the study, several aspects need consideration.

First, my major criticism is that the manuscript needs self-criticism about its limitations. It would benefit from a self-criticism about the small sample sizes for several species (large sample sizes were highly biased to a few species), or some potential limitations about the methods. For example, by using brood patch as an index of breeding activity all species in which only one sex (usually the female) develop a brood patch, on average 50% of the individuals of that species would be wrongly assigned as not breeding. Also, all migratory individuals will necessarily not be in breeding condition at the wintering sites. Explain how these shortcomings might have affected you results. Furthermore, ectoparasitism was classified as either yes or no, but most birds in any region will have a very low load of ectoparasites, thus with potential low effect on hosts. A better approach would be to use ectoparasitism intensity either as a continuous variable or as in classes (ex., zero, small load, heavy load). Thus, ectoparasite presence/absence might not have been a good method, and this should also be addressed in the Discussion.

Second, the final argument in the Discussion (lines 422-426) is too strong considering the limitations above. Also, it is contrary to the several studies cited on lines 399-402, and in the third paragraph of the Introduction. At the most, this argument could be used for this study site.

Further comments:

1)	The title has double meaning as written since temperature' can be interpreted either as host temperature or environmental temperature. Also, restinga, though a vegetation typical of the Atlantic Forest biome, is not a true rainforest. It would be more appropriate to use something like "Brazilian Atlantic xeric forest".
2)	Line 82. It is more common to use nest type (open-cup vs. closed) than nest shape.
3)	Line 89. “…, for example, some …” needs rewriting.
4)	Line 100. “Moreover, seasons can also influence the prevalence in birds concerning reproduction”, is awkward and needs rewriting.
5)	Lines 100-102. “Moreover, seasons can also influence the prevalence in birds concerning to reproduction, since during bird’s breeding season may be an increase in adult’s susceptibility to infections, due to changes in its behavior and immunity”. This assertion needs a reference.
6)	Lines 109-116. In this paragraph, insert the 'area of Restinga' in the beginning of the paragraph, or add a new sentence at the end informing the reader that the whole study was conducted at a Restinga site.
7)	Lines 136-142. Invert the order of this paragraph with the next.
8)	Line 140. For birds it is much more common to use “banding” than “tagging”. Same in line 151.
9)	Line 152. Provide an explanation of how you evaluated birds for age (adult or young).
10)	 Line 153. Explain which stage of the brood patch (full patch?) was considered as breeding or not breeding condition.
11)	 Line 204. It seems that you refer to species and not population specific traits.
12)	 Line 206. Use only open-cup x closed.
13)	 Lines 210-212. Explain how you classified the died of each species. Did you follow HBW or Wilman et al. (2014)? How did you access the number of food items? It is unclear as written.
14)	 Lines 214-216. This part is unclear. Did you use mean temperature for which period? The average of the two years of the study? The month of capture? Same for precipitation?
15)	 Lines 251-252. It is unclear why species not infected were excluded. They could be used to test hypotheses 2 and 3.
16)	 Line 262. I would not consider a sample of > 7 as 'well', but simply as better.
17)	 Lines 266-268. The sample sizes of some of these species do not match the numbers in the tables (Table 1, Table 2, Table S1).
18)	 Line 275. You removed individuals with double infections. Justify the reason for the exclusion of these individuals.
19)	 Lines 318 and paragraph start at line 344. Consider what an odds-ratio of 0.03 really means. What is its biological meaning?
20)	 Lines 349-350. “Therefore, it is possible that migratory birds are not suitable hosts for Parahaemoproteus lineages with which they geographically overlap during their annual cycle.” Do you have evidence from other similar studies that could provide support for this argument? Provide a reference.
21)	 Lines 387-388. “… that meet that criterion breeding sites, ...” This sentence is incomplete.
22)	 Line 405. “…including the very host-species that …” This is unclear.
23)	Lines 416 and 443. Why mention only Brazil? The manuscript should have an appeal for a broader scientific community, at least neotropical. It would be much more interesting and useful to mention the neotropical region.
24)	 Line 431. It is not clear in the text that this is a high diverse habitat, of if you refer to high diversity of birds or of parasites. If you mention birds, provide some reference.
25)	 Table 2. Check the names of Elaenia albiceps and E. chilensis in the tables. There seems to be some error.


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: HOST MIGRATION AND ENVIRONMENTAL TEMPERATURE INFLUENCE AVIAN HAEMOSPORIDIANS PREVALENCE: A MOLECULAR SURVEY IN A BRAZILIAN ATLANTIC RAINFOREST
Review round: 2
Reviewer: 1

Output:
Basic reporting: I was pleased to read the revised manuscript entitled “Host migration and environmental temperature influence avian haemosporidians prevalence: a molecular survey in a Brazilian Atlantic Rainforest”. I believe that the authors addressed all of the reviewers’ comments regarding the original manuscript in a succinct and transparent way.  I agree that the phylogenetic tree was largely unnecessary, and the authors now better explain the potential limitations of the study. The figures are more clearly presented, and all the numbers now make sense.

I would only suggest that the authors again go through the manuscript with a critical eye for the English writing. I have found several instances that I would recommend for correction (below I present the corrected version of the sentences). However, I think that there are still sentences that I may have missed that could be improved.

Title- avian haemosporidian prevalence

23-  and are considered a good model to study host-parasite interactions.

30- and parasite lineages

41- at a closer scale

46- vector-borne

49- Valkiūnas (throughout the manuscript)

61- to the decline

95- Moreover, as seasonality

95- associated with

102- parasite prevalence

210- An overdispersion test

228- findings and allow us to assess the discrepancy between the number of positive samples…

231- Among more highly-captured species…

382- these studies’ results

390- infections only at the community level..
Experimental design: no comment
Validity of the findings: no comment
Additional comments: This is a nice study from a very unusual region of Brazil, and I believe the data will be useful to future scientists studying birds and parasites in relation to climate change.


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: COMPARATIVE ANATOMICAL AND TRANSCRIPTOMIC ANALYSES OF THE COLOR VARIATION OF LEAVES IN AQUILARIA SINENSIS
Review round: 1
Reviewer: 1

Output:
Basic reporting: 1. The authors very well explain the importance of the substance obtained from the endangered tree Aquilaria sinensis. However, very little information is provided on the origin or the region where these two different phenotypes are found and what are the environmental variations - this might give a better understanding of the origin of the new phenotypes and possibly what conditions led to this variation.
2. The authors need some proofreading to correct a few grammatical corrections like in line 46,50,52, 409.

3. Also- in Fig2, the authors mentioned ‘Furthermore, the upper epidermis 193  cell thickness in LGS is significantly smaller than LNS, while the difference of lower epidermis 194  cell between LGS and LNS was not significant ‘ there is some discrepancy in the legend and the figure in fig 2H, 2I (UE vs LE).

4. Could the fig 5A be consolidated anyhow to better represent it? Maybe it can be represented in 2 figures for Stem and leaves separately for all the genes?
Experimental design: 1. What kind of tests were performed to note the statistical differences between the groups in fig 2 and fig 3 is not mentioned in the methods.

2. The authors mention that 65Gb odd data was generated from 12 samples that were then assembled using Trinity program to perform a de novo assembly. The average transcript length is shown to be 1418, which very well falls under good transcriptome construction threshold. It might also be worth while to include some statistics of the assembly and the data (e.g concordant/discordant pair/proper pairs, basic config statistics, preprocessing performed -trimmomatic etc)

3. The authors use DESeq for the analysis of differentially expressed genes between the 2 groups. As a QC, if the authors can provide some figure (e.g PCA plot,  heatmap) to evaluate how well the samples from same group are clustered together, it will give a good sense of the variability amongst samples based on transcriptomic profiles.

4. In fig 4F and 5B,C - it will also be intuitive to show the expression across the replicates for the 4 groups (LGS,LNS,SGS,SNS) to see the consistency aeross samples within same groups rather than only showing average for each group.
Validity of the findings: 1. Although the study dwells on an unexplored aspect of differences in the two phenotypes of an endangered tree due to anatomical and transcriptomic differences, it is not very clear how these findings are relevant to the understand or the differences in survival of the tree or production of agarwood between the two phenotypes

2. The phylogenetic analysis does group the GS and NS with the other Aquilaria sinensis and distinct from Gonystylus bancanus  based on  ITS2+trnL-trnF. However if we observe the phylogenetic tree the 3 GS are under one branch, however NS2 and NS1 are not. Is this concerning. Is there a possible biological explanation for this?

3.The differences in the anatomy are explained with very high resolution and clear images. But it might be lacking the potential effects of those differences observed. It might be worth discussing how could these differences affect the production of agarwood or survival of the tree

4. The finding of the DEG in LGs vs LNS and SGS vs. SNS, the genes related to photosynthesis is expected due to some compensatory mechanisms to synthesis light owing to their less chlorophyll rich area. However, the pathogenesis related genes up regulation in the SGS and LGS is very interesting. Is there any evidence to the these Golden Vein trees dying earlier than a normal vein tress as suggested by the authors in line 409 ‘the development of most of the plants infected with pathogens showing yellow vein symptoms would be influenced and sometimes the plants would die finally‘
Additional comments: Overall the authors have studied the molecular and the anatomical differences in the normal and golden vein phenotype of  Aquilaria sinensis. It is very interesting to study this particular phenotype in a tree that is considered ‘endangered’ and is a producer of a very valued substance ‘agarwood’.


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: COMPARATIVE ANATOMICAL AND TRANSCRIPTOMIC ANALYSES OF THE COLOR VARIATION OF LEAVES IN AQUILARIA SINENSIS
Review round: 1
Reviewer: 2

Output:
Basic reporting: 1) Language
(1)The English language is clear and unambiguous. Pay attention to some mistakes of syntax. Lines 115, publishing paper.
2) Background
(1) Your introduction needs more detail about the study of color variegation, especially advances in molecular biology.
3) Article structure, figures, tables. Raw data shared
(1) Figure 2. Where is idioblasts? Please show it in figure legend.
(2) Figure 5. The number order in paper is incorrect.
Experimental design: 1) Original primary research within Aims and Scope of the journal.
Yes
2) Research question well defined, relevant & meaningful.
Yes
3) Rigorous investigation performed to a high technical & ethical standard.
Yes, but need more experiments to support the conclusion. The content of chlorophyll and carotenoid? The ultrastructure of chloroplast?
4) Methods described with sufficient detail & information to replicate.
(1) The method of DNA extraction. Line 111, please show the company name of this manufacturer belongs to.
(2) The method of quantitative real-time PCR. Lines 161-162, check the procedure of RT-PCR.
Validity of the findings: 1) All underlying data have been provided; they are robust, statistically sound, & controlled
Yse
2) Conclusions are well stated, linked to original research question & limited to supporting results.
Yse
Additional comments: 1) Lines 178-180. Please show the result of gel electrophoregram in supplemental figure.
2) Line 220. Each with three biological duplicates or technical duplicates?
3) Lines 238. What is the meaning of DE? The authors need to make sure that they use the full names of all the factors before switching to abbreviations.
4) The species names and gene names should be italic. Please go through the manuscript including references and ensure that all species and gene names are italicized.


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: COMPARATIVE ANATOMICAL AND TRANSCRIPTOMIC ANALYSES OF THE COLOR VARIATION OF LEAVES IN AQUILARIA SINENSIS
Review round: 2
Reviewer: 1

Output:
Basic reporting: NA
Experimental design: NA
Validity of the findings: NA
Additional comments: All the concerns have been addressed.


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: COMPARATIVE ANATOMICAL AND TRANSCRIPTOMIC ANALYSES OF THE COLOR VARIATION OF LEAVES IN AQUILARIA SINENSIS
Review round: 2
Reviewer: 2

Output:
Basic reporting: 1.	Figure 2: it would be interesting to specify the P Value that graphs have with a single asterisk.
Not all the parts names of the leaves look good. Change color, size, bold… Especially figure C.
2.	Figure 5B: A and B are the same sentence. I understand that in B it should be SGS and SNS. The legend in Figure B also contains the mistake.
3.	Figure 6: Better specify the letter E and F. It will help to understand the figure more easily.
4.	Table S5: change de novo  de novo
5.	Littles mistakes:
a.	Line 21 – 22 and 206: different size of letter
b.	Line 135: change trnL  trnF
c.	 Line 196, 219 – 224, 262, 416 – 417: highlight the figures in bold
d.	Line 212: change abd  and
6.	In the paragraph line 264 “Photosynthesis related genes….” Figure 5C is not referenced.
Experimental design: 1.	Line 114- 116: the procedure that has been used for sample collection is not well understood. It would be interesting to specify how the collection was, in number of leaves or stems that were collected, mixed ... The experimental design that has been carried out is very important.
2.	Line 368: The authors select 22 genes for validation but in graph 6D they are 23. Please revised.
3.	Line 371: the authors explain “our data revealed that most of the gene expression data in qRT-PCR matched well with transcriptome results”. The authors could provide some figure o table (e.g correlation graph or similar) to evaluate this.
Validity of the findings: No comments to highlight in this section.
Additional comments: The authors have compared two different types of phenotypes in Aquilaria sinensis at the anatomical and transcriptomic level. It is a very complete and interesting work, in which they provide a large amount of data about these two phenotypes.


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: COMPARATIVE ANATOMICAL AND TRANSCRIPTOMIC ANALYSES OF THE COLOR VARIATION OF LEAVES IN AQUILARIA SINENSIS
Review round: 2
Reviewer: 3

Output:
Basic reporting: The manuscript is written in clear, professional English. The introduction section sets up the premise of the paper very well, and provides enough context about the developments in the field and how this paper serves as an advance. The extensive anatomical comparison of leaves and stems is particularly commendable and shows the comprehensive nature of this study that explores the color variation across two types of leaf samples Aquilaria sinensis
Experimental design: The research question is well defined and meaningful within the scope of the journal.
The investigation performed is rigorous and meets a high level of technical standard.
One minor suggestion is to clearly state which background gene sets are being for GO/KEGG pathway enrichment - it looks like this would be the list of 18,786 genes with functional annotation but should be mentioned.
Also, the in Figure 6E,F the labels for the two y-axes bars should be mentioned, and a short description of what they are should be included in the caption
Validity of the findings: Conclusions are stated clearly and are supported by the results reported by the authors
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: COMPARATIVE ANATOMICAL AND TRANSCRIPTOMIC ANALYSES OF THE COLOR VARIATION OF LEAVES IN AQUILARIA SINENSIS
Review round: 3
Reviewer: 1

Output:
Basic reporting: Thanks for your answer.
No comments to highlight in this section.
Experimental design: Figure S5: it would be interesting to better explain the bar graphs for a better understanding
Validity of the findings: No comments to highlight in this section.
Additional comments: Thanks for anwers.


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: COMPARATIVE ANATOMICAL AND TRANSCRIPTOMIC ANALYSES OF THE COLOR VARIATION OF LEAVES IN AQUILARIA SINENSIS
Review round: 3
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: CONTINUED NUCLEIC ACID TESTS FOR SARS-COV-2 FOLLOWING DISCHARGE OF PATIENTS WITH COVID-19 IN LU’AN, CHINA
Review round: 1
Reviewer: 1

Output:
Basic reporting: 1. An essential advice for authors is to consult a professional native English-speaking editing service for assistance with the manuscript, as there are several parts through it that should be carefully reviewed. Unfortunately, the manuscript is plagued with an undesirable number of spelling and grammatical errors, and poor punctuation that should be solved to improve the paper. As a consequence, some parts of the manuscript are hard to follow. Authors should understand that the reviewers´ function is to evaluate the quality and accuracy of the research presented, and provide some feedback to improve the clarity, transparency, accuracy, and utility of potential papers, not to serve as English spelling and grammar correctors.

2. Unfortunately, the study lacks a solid and clear theoretical/conceptual framework, which makes it hard to justify the study.

3. The purpose of the introduction is to orient the reader and create an interest in the study. In this manuscript, however, the introduction is too vague and makes it hard to justify it. From my perspective, there is no coherence and cohesion between the introduction and the results and conclusions to make it easier for readers to follow the  authors´ arguments. Thus, authors are invited to provide a more extensive and clear introduction highlighting the importance of running a study like this. Authors should aim for a logical order that helps readers make sense of the introductory information.
Experimental design: 1. The research question is ambiguous. Furthermore, the rationale for the study is not clear at all. The authors are kindly invited to provide a clear statement about the need and relevance of this study.

2.  I would like to know if the research protocols used in this study were reviewed and accepted by an independent committee before initiation. The authors should mention if informed consent from research participants were given.

3. The methods need to be better described. The Methods section should give readers enough information in such a form that they can repeat the experiments. In this manuscript, several methods are unclear. Thus, the authors mention that the Sun Yat-Sen University manufactured the RT-PCR kit used, but no information about the target sequence or amplification conditions is given. In this way, interested readers may found it hard to follow the same procedure.
Validity of the findings: 1. The description of the results (line 80-94) is too confusing. It is hard to follow, and data are hard to follow. The authors are invited to provide a table resuming this information.

2. From my perspective, the weakest parts of the study are the discussions and conclusions. This may be due to the lack of clarity on the results obtained.

3. The discussion is not a real discussion. The purpose of the discussion is to interpret and describe the significance of the findings in light of what has been previously published about the research problem being investigated. However, in this study, the authors cite only three studies to "discuss" their findings. Thus, the discussion lacks depth and seems biased to favor what the authors assume.
Additional comments: This could be a very interesting study if authors prepare a better manuscript. It is important to provide a scientifically supported discussion and give more clarity on methodology and results description.

I would also like the authors to look for some professional help from a native English-speaking editing service. In its actual form, several parts of the manuscript are too hard to follow.


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: CONTINUED NUCLEIC ACID TESTS FOR SARS-COV-2 FOLLOWING DISCHARGE OF PATIENTS WITH COVID-19 IN LU’AN, CHINA
Review round: 1
Reviewer: 2

Output:
Basic reporting: Dear Authors,

The English language of your background section (lines 19 and 20) should be improved. This improvement can help an international audience clearly understand the message that you are trying to convey. For example, "four laboratory confirmed COVID-19 cases that were confirmed to be positive for SARS-CoV-2 by RT-PCR following discharges." You can improve this sentence by writing, "during their follow-up, four discharged COVID-19 individuals/patients were RT-PCR retested positive again for SARS-CoV-2." This is similar to this article: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7276353/#r2. Also, according to the PeerJ's Standard Sections (https://peerj.com/about/author-instructions/#standard-sections), subheadings should be followed by a period (e.g., Background.). Your subheadings were followed by colons (e.g., Background:, Methods:, Results:).

The English Language of your methods section (lines 22-25) should be improved as well. You can improve these sentences (lines 22-24) by writing, "We conducted RT-PCR tests on sputum, throat swab, fecal or anal swab and urine samples collected from 67 COVID-19 patients/individuals following their discharge in Lu'an Municipality, China (is this a hospital?). These samples were collected on the 7th and 14th day following their discharge." Lines 24 and 25 or the sentence, "In case the positive interval was 24 h, the samples were collected again until they were confirmed as negative for 2 times.", should be improved. If you are collecting samples on the 7th and 14th day, can you clarify the 24 hours period? Is that if a patient/individual tested positive on the 7th or 14th day, you waited 24 hours before testing them again?

The English of your results section (lines 26-32) should be improved as well.

You should have cited the following articles/reports:

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7276353/#r2

https://ophrp.org/journal/view.php?doi=10.24171/j.phrp.2020.11.3.02

https://www.cidrap.umn.edu/news-perspective/2020/05/wha-passes-pandemic-probe-resolution-korea-clarifies-reinfection-reports

https://www.frontiersin.org/articles/10.3389/fcimb.2020.00445/full

https://www.cebm.net/covid-19/infectious-positive-pcr-test-result-covid-19/

https://www.medrxiv.org/content/10.1101/2020.08.04.20167932v3 (this was published after you submitted the paper)
Experimental design: Dear Authors,

The experimental design of your manuscript should be changed. A recent systematic review by the Centre for Evidence-Based Medicine (CEBM) (https://www.cebm.net/covid-19/infectious-positive-pcr-test-result-covid-19/ ), “Are you infectious if you have a positive PCR test results for COVID-19?”, highlights the issue of using, only, PCR results to make clinical diagnosis or decisions. Jefferson et al. stated that, “PCR detection of viruses is helpful so long as its accuracy can be understood: it offers the capacity to detect RNA in minute quantities, but whether that RNA represents infectious virus may not be clear. During our Open Evidence Review of oral-fecal transmission of Covid-19, we noticed how few studies had attempted or reported culturing live SARS-CoV-2 virus from human samples.” Also, the BBC (https://www.bbc.com/news/health-54000629) wrote an article, yesterday (September 5th), about this systematic review. Furthermore, back in May of this year, “South Korean researchers revealed that discharged patients who test positive for COVID-19 again don't appear to be contagious.” This was reported by the Center for Infectious Diseases Research and Policy (CIDRAP) (https://www.cidrap.umn.edu/news-perspective/2020/05/wha-passes-pandemic-probe-resolution-korea-clarifies-reinfection-reports). The CIDRAP’s report explained that, “South Korea's investigation into recovered patients who test positive for COVID-19 again found no live virus in the patients they examined, suggesting no risk of passing the virus to another person and that the patients were likely shedding noninfectious or dead virus particles, the Korea Centers for Disease Control (KCDC) said today.” This led to a change in the KCDC’s discharge protocols (https://www.cdc.go.kr/board/board.es?mid=a30402000000&bid=0030&act=view&list_no=367267&nPage=1). According to the new KCDC’s discharge protocols, “no additional tests are required for cases that have been discharged from isolation.” Finally, their discharge protocol now states that, “the terminology for referring to such cases will be changed from “re-positive” to “PCR re-detected after discharge from isolation”.”

Your experimental design can be improved by conducting viral cell culture tests along with RT-PCR. This would be more useful to reexamine the current discharge criteria in Lu’an Municipality, china.
Validity of the findings: Dear Authors,

I think your results should have been validated using viral cell culture. Please use the paragraph bellow as a reference to improve your experimental design.

"Conclusion Prospective routine testing of reference and culture specimens are necessary for each country involved in the pandemic to establish the usefulness and reliability of PCR for Covid-19 and its relation to patients factors. Infectivity is related to the date of onset of symptoms and cycle threshold level. A binary Yes/No approach to the interpretation RT-PCR unvalidated against viral culture will result in false positives with segregation of large numbers of people who are no longer infectious and hence not a threat to public health."

https://www.medrxiv.org/content/10.1101/2020.08.04.20167932v3
Additional comments: Dear Authors,

Your manuscript is relevant to the current COVID-19 global pandemic and is in line with other publications related to COVID-19 patients testing positive after discharge. This reviewer is conscious of the tremendous amount of work and effort that went into the manuscript. Maybe you do not have access to a biosafety level 3 (BSL 3) laboratory to conduct viral cell culture experiments. However, viral cell culture experiments are important in order to accurately make clinical decisions (e.g., change discharge protocols). These decisions are crucial not only to save lives, but also to save resources.