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: LONG-TERM CONTINUOUSLY MONOCROPPED PEANUT SIGNIFICANTLY CHANGED THE ABUNDANCE AND COMPOSITION OF SOIL BACTERIAL COMMUNITIES
Review round: 2
Reviewer: 2

Output:
Basic reporting: I still strongly suggest the language be revised by a native English speaker. For instance，in line 138,  "distance" is very confusing, do the authors mean "depth" ?
Experimental design: It is still not possible to figure out from which rhizocompartment the soil was collected? rhizosphere? rhizoplane or bulk soil?
Validity of the findings: It is still hard to figure out a clear conclusion of the manuscript mainly due to the language issue. For example, the term "simplification of bacterial community", which has been described as the main reason for the peanut soil sickness, is very confusing.
Additional comments: Most of my comments in the first round review have not NOT 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: LONG-TERM CONTINUOUSLY MONOCROPPED PEANUT SIGNIFICANTLY CHANGED THE ABUNDANCE AND COMPOSITION OF SOIL BACTERIAL COMMUNITIES
Review round: 3
Reviewer: 1

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


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

Input:
Title: TWO SPECIES OF THORACOSTOMOPSIDAE (NEMATODA: ENOPLIDA) FROM JEJU ISLAND, SOUTH KOREA
Review round: 1
Reviewer: 1

Output:
Basic reporting: Manuscript by Jeong, Tchesunov and Lee is devoted to description of one new species and one known species nematodes from the family Thoracostomopsidae. Important parts of the study are the revision of the genus Enoploides and identification key.
The English language should be improved to ensure that an international audience can clearly understand your text.
The list of reference is quite full, but there are some reference missed:
Ditlevsen 1919; Wieser & Hopper, 1957.
Introduction part is clear and explains well the aim of the study.
Drawings look like drafts and must be improved for publishing. The presence of SEM photo is a advantage for the paper. But light microscopical photo are highly recommended. It would not take a lot of time for authors as fas as they have made voucher photo for sequenced specimens.
Experimental design: Although it is very important to accumulate the data on DNA of free-living nematodes in GenBank, the size of 18S rRNA site amplified is only about 300 bp. Unfortunately, It is very short site for this gene. Such a short site is not enough to make any reliable conclusions on phylogeny of the group discussed. It is better to omit the part concerning phylogenetic tree and its discussion.
The descriptions are very detailed. Usually description of nematodes are made in telegraph style. For example, instead of "four shorter cephalic setae sharing one crown" (line 430) better use "six outer labial setae and four cephalic setae in one crown".
Line 438 "cervical somatic setae distributed along  the body in singlets". Cervical setae can not be scattered along the body as fas as they are situated exclusively in cervical region. There is no need using term "in singlets" if the single setae scattered along the body. Better say "Cervical setae irregularly scattered along the cervical region"
Line 438 Amphid not observed. May be more careful examination of slides and SEM stubs will help?
Line 440 What does it means "pharynx annulated"?
Line 444 "82 mkm long above cloacal opening" Do you mean 82 mkm above cloacal opening?
Line 455 "Reproductive system didelhpic amphidelphic flexed inwards" the sentence is not clear. Please, clarify it in drawing as well. It does not look amphidelphic in drawing.
Key to valid species of Enoploides is very important and careful part of the manuscript. Line 548-549 What is post-anal cuticular element? There is no such elements in original drawings. May be the additional picture of this element would be helpful for manuscript readers.

The same comment to the secription of the second species, please, steak to telegraf style.
Line 607-608 "Base of mandibles widening near the base" Do you mean base of the base? Or just base?
Line 616. "Caudal glands protruding anterior to the anus" Better say "caudal glands in pre-anal position"
Validity of the findings: Conclusions are well stated, all underlying data have been provided.
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: TWO SPECIES OF THORACOSTOMOPSIDAE (NEMATODA: ENOPLIDA) FROM JEJU ISLAND, SOUTH KOREA
Review round: 1
Reviewer: 2

Output:
Basic reporting: .
Experimental design: .
Validity of the findings: .
Additional comments: See attachment


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: TWO SPECIES OF THORACOSTOMOPSIDAE (NEMATODA: ENOPLIDA) FROM JEJU ISLAND, SOUTH KOREA
Review round: 1
Reviewer: 3

Output:
Basic reporting: Title: Two species of Thoracostomopsidae (Nematoda: Enoplida) from Jeju Island, South Korea

Reviewer #1
General Comments
In this study, the authors described a new species of Enoploides koreanus sp. nov. and redescribed Epacanthion hirsutum Shi & Xu 2016, both belonging to the family Thoracostomopsidae (Nematoda: Enoplida). Some genetic data of the two species were also reported. The genus Enoploides was reviewed based on references and a key to species was provided. The illustrations and SEM micrographs are of good quality. However, there are a number of serious flaws: (i) The name of the new species is not correct, because the generic name Enoploides is a feminine gender, but specific name “-koreanus” is masculine gender， which are conflicted. (ii) The 18S rDNA with very slow evolutionary rate is not suitable for species identification of nematodes. So Table 7 has no any scientific significance, that could not support your specimen collected from Jeju Island to be congeneric with Epacanthion hirsutum. (iii) The method for phylogenetic analysis is very rough (lacking of details). Which group was chosen as outgroup? Most species of Enoploides with 18S sequence data available in GenBank, i.e. Enoploides delamarei, Enoploides longispiculosus, Enoploides sp. 161216.19 etc. have not been included in the phylogeny. (iv) the authors did not examine any type or identified voucher specimens of Enoploides species for revision of this genus, thus the reliability of results is weak. I strongly suggest the authors check some type or identified voucher specimens to confirm the taxonimical status of some species of Enoploides with debated validity. (v) the English of manuscript needs much improvement. The text (i.e. abstract, Introduction, description) is wordy and has some grammatical errors. It would benefit from a revision made by a native speaker.

Specific comments
1.	Abstract: the present Introduction is rather wordy. Some sentences have obvious grammatical errors. Please rewrite it.
2.	Please do not use the abbreviation in abstract, give the meaning of the letter ”b”.
3.	Introduction, Line 81-82: The aim of this study was to review and revise the genus Enoploides while reporting a new species....you did not revise the genus Enoploides?
4.	Materials and Methods: Which group was chosen as outgroup for the phylogeny? How many bootstrap replications did you test?
Experimental design: (i) The authors used the 18S rDNA as genetic marker for speciese identification in the present study. However, the 18S rDNA with very slow evolutionary rate is not suitable for species identification of nematodes. So the result of Table 7 could not support the specimen collected from Jeju Island to be congeneric with Epacanthion hirsutum. (ii) The method for phylogenetic analysis is very rough (lacking of details). Which group was chosen as outgroup? Most species of Enoploides with 18S sequence data available in GenBank, i.e. Enoploides delamarei, Enoploides longispiculosus, Enoploides sp. 161216.19 etc. have not been included in the phylogeny.
Validity of the findings: (i) the authors did not examine any type or identified voucher specimens of Enoploides species for revision of this genus, thus the reliability of results is weak. I strongly suggest the authors check some type or identified voucher specimens to confirm the taxonimical status of some species of Enoploides with debated validity.
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: TWO SPECIES OF THORACOSTOMOPSIDAE (NEMATODA: ENOPLIDA) FROM JEJU ISLAND, SOUTH KOREA
Review round: 1
Reviewer: 4

Output:
Basic reporting: I found the paper very orginal, well written and structured. I think that this type of taxonomical papers is really necessary because it is complete and increases our knowledge on the diversity of an important nematode family. In particular, I have appreciated that the authors analysed their samples with the aid of several tecniques (i.e. SEM, light microscopy and molecular approach) giving a complete description of the new species. Furthermore, I think that the new indentification key, theupdate list of valid species and new molecular insights will be important for the taxonomists of this phylum. Thus, I suggest the pubblication of this paper after minor revisions.
I reported in the file some minor changes that I suggest to the authors. The main ones are a doubt in the identification key and the suggestion to add further details in the SEM picture of the male of Enoploides koreanus
Experimental design: The approaches used are fully in agreement with the current taxonomical standards.
Validity of the findings: It is one of the most complete taxonomical paper that I have reviewed and I think that it will have a high relevance in the field.
Additional comments: lines 307-308; 312-313; 337-338; 342-343; 363-364; 368-369; 373-374; 390-391. I suggest to revise the english of this sentences;
line 411 Please add the total number of specimens analysed
line 530 Please check this point also considering the point 3 (see line 527) in which you stated that pre anal supplements were present ;
line 643: Change as follows "primer sets utilized in the present study."
line 673  delete italic in the following word "Mawson"
Fig 1 is it possible to add a drawing with the spicule and gubernaculum details?
Fig 3 B in my opinion it is quite difficult to see setae. Could you add a detail increasing the magnification?
Fig 3 C Also in this case I suggest a detail of the tip


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: TWO SPECIES OF THORACOSTOMOPSIDAE (NEMATODA: ENOPLIDA) FROM JEJU ISLAND, SOUTH KOREA
Review round: 2
Reviewer: 1

Output:
Basic reporting: I am satisfied with revision of manuscript. It could be accepted for publication.
Experimental design: Methods described with sufficient detail & information to replicate.
Validity of the findings: All underlying data have been provided; they are robust, statistically sound, & controlled.
Additional comments: Authors said that they added Phillips et al., 2016 to line 106, but I have not found it. It is also absent in reference list.


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: TWO SPECIES OF THORACOSTOMOPSIDAE (NEMATODA: ENOPLIDA) FROM JEJU ISLAND, SOUTH KOREA
Review round: 2
Reviewer: 2

Output:
Basic reporting: The language of Ms is clear now. The introduction is good enough.
Experimental design: The research question has been well solved. The methods used in the study is suitable and powerful.
Validity of the findings: The authors described a new species of Enoploides koreanus sp. nov. and redescribed Epacanthion hirsutum Shi & Xu 2016, both belonging to the family Thoracostomopsidae (Nematoda: Enoplida) and provided important morphological and genetic data for this group.
Additional comments: This revised version of manuscript has been significantly improved. All of my recommendations have been complied. I have 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: TWO SPECIES OF THORACOSTOMOPSIDAE (NEMATODA: ENOPLIDA) FROM JEJU ISLAND, SOUTH KOREA
Review round: 3
Reviewer: 1

Output:
Basic reporting: See comments in the annotated PDF.
Experimental design: See comments in the annotated PDF.
Validity of the findings: See comments in the annotated PDF.
Additional comments: See comments in the annotated PDF.


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: KNOCKDOWN OF CDCA8 INHIBITS THE PROLIFERATION AND ENHANCES THE APOPTOSIS OF BLADDER CANCER CELLS
Review round: 1
Reviewer: 1

Output:
Basic reporting: No comment.
Experimental design: No comment.
Validity of the findings: No comment.
Additional comments: The present study has found an oncogene called CDCA8 (cell division cycle associated 8) which could promote proliferation and inhibit apoptosis in bladder cancer. They used bioinformatics analysis as a premise, combing with the in vitro experiment to complete the study. The results are convincing and make some sense, but I still have some suggestions to improve the article:
1.	The length of the introduction is too short. Please search more reference to support your idea.
2.	The phrase “RT-qPCR assay” is wrong.
3.	In spite of GSE13507, more datasets such as TCGA-BLCA, GSE32584 and GSE48075 etc. should be analyzed to see whether CDCA8 is upregulated and correlated with poor prognosis in all this datasets.
4.	The overexpression experiment should be added to validate the role of CDCA8 in bladder cancer.
5.	The western blot of marker proteins such as CCND1, caspase etc. should be 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: KNOCKDOWN OF CDCA8 INHIBITS THE PROLIFERATION AND ENHANCES THE APOPTOSIS OF BLADDER CANCER CELLS
Review round: 1
Reviewer: 2

Output:
Basic reporting: 1. The sentence “The expression of … was knocked down” is not accurate (line 41, for example). It is rather the gene that is knocked down and not its expression
2. Sentences at lines 67, 82 have no reference.
3. Line 77. When referring to the authors of an article by name “Dai et al.” the intext citation that follows should be only the year in parentheses to avoid repetition, unless the journal has explicit format that cover this case. Alternatively, the full intext citation can be moved to the end of the sentence.
4. Line 89. “... analyzed by bioinformatics.” The authors should consider being more specific about with methods and tools were used to analyze the correlation.
5. The GEO datasets were wrongly referred to in the text as “high throughput sequencing datasets ” (line 87) and “chip sequencing data” (line 94). The four datasets were generated using microarrays!
6. The method to obtain the datasets and the form of the data (raw/processed) were not mentioned.
7. The transformation of gene probe ID to gene symbol (line 99) is rarely as simple as that. How did the authors map probes from different platforms to gene symbols? How did the authors deal with probes mapping to multiple genes and genes represented by multiple probes? Also, “international standard gene names” doesn’t refer to any specific naming convention/standard.
8. Line 119. The term WB was not mentioned in full.
9. The calculation of the delta delta Ct value was described as subtracting the mean delta Ct of control from each sample separately. Some argue that the means of two groups should be used. Please justify or cite the source. Also the calculation of the error term was not described.
10. The term “dataset” and “data set” were both used in the text. Similarly, the terms “knockdown” and “knockout” were used interchangeably. Please revise for consistency.
11. Figure 1. The x-axis doesn't start from 0 and the figure legend is very brief.
12. Figure 3, 6-8 legend are very brief.
Experimental design: 1. The authors chose four independent datasets of normal and bladder cancer tissue to visualize the expression of CDCA8 and associate it with clinical characteristics and outcomes. The choice of these particular datasets was not justified. Large studies such as those of the cancer genome atlas can be a better choice if the main goal is to establish links between the expression and patient metadata.
2. The correlation of gene expression with clinical characteristics was not formally analyzed. Although figure 2 provide useful visualization of the expression in different groups, to claim significant correlations, they should be tested in regression models.
3. Gene and protein markers were not used to study the effect of the knockdown of CDCA8 on cell cycle or growth.
Validity of the findings: 1. The visualization of the expression of CDCA8 in multiple bladder cancer microarrays datasets is valid. However, the claim that this gene correlates with the tumor, stage, grade and age was not well established. The use of multiple regression model is required to establish the associations.
2. The functional study of the CDCA8 knockdown bladder cancer cell lines is suggestive. However, the mechanism by with this protein might be working was not fully investigated. The cell cycle analysis is useful but doesn’t justify the strong claims. First, analysis of key gene and protein markers of proliferation, migration, invasion, apoptosis and cell cycle is missing. Second, pattern of cell cycle regulation is in one cell line is opposite to the other. At least, this difference should be addressed in the discussion.
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: KNOCKDOWN OF CDCA8 INHIBITS THE PROLIFERATION AND ENHANCES THE APOPTOSIS OF BLADDER CANCER CELLS
Review round: 1
Reviewer: 3

Output:
Basic reporting: .
Experimental design: .
Validity of the findings: .
Additional comments: Authors of the present study investigated the  correlation between the expression of
CDCA8 in bladder cancer and clinical variables as well as  the role of CDCA8 in bladder cancer cells in vitro. The aim of the study is original. However, some revisions are needed in order to improve the overall quality of the manuscript.
Methodology: I suggest to improve the methodology by analyzing, in a multivariable model including also clinical variables, the predictors of survival.
Discussion: I suggest  to improve the discussion about  the significant correlations found between CDCA8 expression and some clinical 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: KNOCKDOWN OF CDCA8 INHIBITS THE PROLIFERATION AND ENHANCES THE APOPTOSIS OF BLADDER CANCER CELLS
Review round: 1
Reviewer: 4

Output:
Basic reporting: The article is written in a clear and concise manner. The introduction has depth with good background information. The authors have, however, skipped referencing a paper - Bi et al, 2018 “CDCA8 expression and its clinical relevance in patients with bladder cancer” which is very closely related to this article. It would be good for them to reference this article and potentially in the discussion compare and contrast their results with this article. The discussion is a little repetitive and may be overstating the use of CDCA8 as a prognostic marker as high expression is seen in many patients with good prognosis. Good detail in the materials and methods regarding the experimental techniques; but more detail is necessary regarding computational methods/bioinformatics with respect to the different analysis performed. It would be worthwhile to compress the figures from 9 to 4 to allow for a more streamlined flow of the paper and the data. For instance, the migration and transwell assays can easily be included as the same figure and similarly for the comparison of levels of CDCA8 and clincal features and survival curves. More detail can be added to the figure legends of figure 5,6,7,8,9.  Some editing for language and grammatical errors would be useful before the publishing of this manuscript.
Experimental design: The experimental design is suitable to the goal that the authors have outlined and the article is within the scope of the journal. The authors have described a knowledge gap in the field by stating that role of CDCA8 in bladder cancer in unknown. By stating the role of CDCA8 in tumorigenesis in other cancers, the authors justify their preliminary notion that CDCA8 might have an important role to play in bladder cancer as well. Using bioinformatic analysis of published microarray data as well with comparison with clinical data of those samples, the authors have identified that CDCA8 is overexpressed in bladder cancer cells relative to normal cells. Further, they used in vitro assays and identified that CDCA8 knockdown in the bladder cancer cells results in the reduced proliferation, migration, invasion, cell cycle arrest and increased apoptosis. While the authors have performed these in vitro studies using two different cell lines, no information is provided regarding the number of biological replicates done and whether the quantification is representation of multiple replicates. This would be useful to include in the final version of the article.
Validity of the findings: The authors aim to identify the role of CDCA8 in bladder cancer. By using a two pronged approach of bioinformatics and in vitro assays, they provide good evidence to support their hypothesis. The following changes/experiments would be useful to strengthen the paper.
1)	In Figure 1, the authors identify that CDCA8 expression is increased in bladder cancer tissue relative to normal tissue. It would be useful to provide more information regarding the studies that were used. It can also be noted that the levels of CDCA8 in normal tissues varies amongst the studies as well as the number of normal tissues used for comparison are very low – a sentence or two of comment regarding this would be useful. Finally, given the availability of large resources now such the TCGA as well as other large RNAseq dataset, it would be good to see corroborative evidence from the analysis of a larger dataset  or justification as to why this was not done. Lastly, comparison of levels of CDCA8 to known markers of bladder cancer as well negative controls (house keeping genes) would provide strength to the study showing the CDCA8 is indeed a marker for bladder cancer.
2)	In Figure 2, the authors use both p=value and sometimes p<value in different sub-figures; this would need to changed to be of the same format in all the figures with a clear NS written in graphs describing non-significant comparisons. Further, the authors claim that the expression of CDCA8 is highly and significantly correlated with a clinical factor – however, no correlation analysis has been performed to justify this claim. I would recommend that the language be amended to “ CDCA8 expression is higher in individuals with high grade tumor when compared to individuals with low grade tumor” etc.
3)	In figure 4, an explanation needs to provide in the methods or results regarding what is driving the GFP expression and how the the CDCA8 knockdown results in a loss of GFP expression. While the mRNA expression and protein expression are convincing regarding the CDCA8 knockdown, the knockdown is not apparent looking at the fluorescence images.
4)	In figure 5, while the effects on CDCA8 knockdown on the growth of these cells is striking, corroborative experiments such as Brdu incorporation etc would be useful to add strength to the claim that these cells have reduced proliferation.
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: KNOCKDOWN OF CDCA8 INHIBITS THE PROLIFERATION AND ENHANCES THE APOPTOSIS OF BLADDER CANCER CELLS
Review round: 2
Reviewer: 1

Output:
Basic reporting: The author have improved their writing.
Experimental design: No comment.
Validity of the findings: No comment
Additional comments: 1. I think the dataset GSE48075 do include patient prognosis information as I have used this dataset in my study. So I also recommend author to make some analysis in GSE48075.
2. I think some markers associated with cell cycle, apoptosis should be done to prove CDCA8 really influence these process.


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: KNOCKDOWN OF CDCA8 INHIBITS THE PROLIFERATION AND ENHANCES THE APOPTOSIS OF BLADDER CANCER CELLS
Review round: 2
Reviewer: 2

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The revised manuscript was improved as a result of:
Using TCGA data to visualize the expression of CDCA8
Performing univariate and multivariate analysis to establish CDCA8 as prognostic factor

The authors chose to not extend the study into the mechanism by which this protein might influence the proliferation or the cell cycle of bladder cancer. While this represents an issue of the rigour of the work, it has been acknowledged in the discussion and the conclusion.

Two points remains to be resolved however. First, no correlation was observed between CDCA8 expression and the patient prognosis. I’d be inclined to further investigate this point. Second, two cell lines had opposing patterns of cell cycle regulation. While this was reported as such, no explanation was provided besides “may be CDCA8 has different effects on the cell cycle.” This might be true, but it basically contradicts the rest of the study. Therefore, observing the desired pattern in the other cell line cannot be taken as supporting evidence.


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: ESTIMATION OF SOIL SALT CONTENT BY COMBINING UAV-BORNE MULTISPECTRAL SENSOR AND MACHINE LEARNING ALGORITHMS
Review round: 1
Reviewer: 1

Output:
Basic reporting: The presentation of this work is well down, especially Abstract part is well written. But Some minor problem need attention.
Line 37: It is recommended not to appear “first”, “Subsequently”, and “finally” in the method description section.
Line 46: The result description keeps the tense consistent and should use the past tense. Such as: “the result showed” corresponding is “had been improved”.
Line 61-64: “the process of salinization……”, this sentence is not native. It is recommended to rewrite it in two sentences.
Line 64 and line 199: “To manage” instead of “in order to manage”, “to find” instead of “in order to find”
Line 81-85: This explanation is taking forever. It is recommended to revise.
Moreover, the authors should pay attention to the uniform format of the article paragraph.
Experimental design: Sample selection and experimental design are satisfactory. I would like the author to explain why not consider RPIQ as a model validation indicator. Because SSC is generally non-normal. “software Pix4D” should include information such as version and company.
Validity of the findings: 1. Data is robust, statistically sound, & controlled.
2. Conclusions are well stated, linked to original research question & limited to supporting results.
3. Speculation is welcome.
Additional comments: 1. “variable selection method” is a better way of putting it than so-called “dimensionality reduction”. VIP, SPA and methods are mainly for variable optimization.
2. Figure 6 is not clear, and the value of the diagonal should be NaN. (B) omit Y-axis.
3. I have not found the relevance of the previously mentioned published indices and SSC.
4. Figure 10, red Y-axis should put near the blue Y-axis.
5. Three machine learning algorithms belong to three fields respectively, for example, RF belongs to ensemble learning, which advantages it plays (except for being appropriate for small samples).


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: ESTIMATION OF SOIL SALT CONTENT BY COMBINING UAV-BORNE MULTISPECTRAL SENSOR AND MACHINE LEARNING ALGORITHMS
Review round: 1
Reviewer: 2

Output:
Basic reporting: no
Experimental design: no
Validity of the findings: no
Additional comments: This paper investigates modeling of soil salt content using UAV-borne multispectral data. Machine learning methods was found to be most useful. The UAV-borne multispectral remote sensing has great potential for estimating SSC in the future; and a suite of optimized spectral indices were proposed to estimate SSC. The topic is very important given the fact that soil salinization is a very important element in sustainable agriculture. This research paper presents an interesting concept toward that front, and interesting to the readers of the journal. I recommend publications with minor revision. summary comments are below:

1.	The contents of the manuscript were too much redundant and should be substantially condensed. The advantages of remote sensing in the Introduction section was common knowledge, and should be drastically cut down.
2.	It is well known that soil moisture affects the spectral reflectance. The existence of such difference needs to be considered in the subsequent studies, and the reasons for such difference can be analyzed in depth.
3.	I noticed a few citations which follow statements that do not accurately reflect the findings of what is being cited (Line 221-222…..). I recommend you review your citations and ensure that the way they are being presented in your paper is accurate.
4.	Why the full-covariates models have less good performances? Can we just imagine the less important covariates as some kind of noises, and these noises compromise the models performances?
5.	The authors should add some related discussion about the estimation mechanism. I think your paper can best be improved by adding the related contents.
6.	Based on what considerations did you determine the threshold values (e.g., 0.7 for GRA and 1.0 for VIP) for band selection methods?
7.	Please specify the detailed information of “Pix4D mapper”
8.	This paper did not present the relationship between the SSC and other spectral indices. It is better to add this part.
9.	Line 423:”CONCLUSION” should be on the next line.
10.	Line 424: “nine estimation models” or “twelve estimation models”, please check it.


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: ESTIMATION OF SOIL SALT CONTENT BY COMBINING UAV-BORNE MULTISPECTRAL SENSOR AND MACHINE LEARNING ALGORITHMS
Review round: 1
Reviewer: 3

Output:
Basic reporting: Line 61, be resulted from or ...is due to...

Line 62, vague, what is the "specific natural conditions"

Line 79, none of these four papers addressed UAV can monitor heavy metals if I am reading the titles of the four references correctly

Line 99, represent soil salinization, not monitor

Line 106, filter the redundant information, not reduce..

Line 141, missing year for Wang et al.
Experimental design: Line 157, uppercase for km2.

Line 158-163, missing references for climate and soil texture

Line 216 and 224, please write out all the variables. And these formulas are complicated, if this is a function in R or other software, readers might not need to see the whole formula since they don't need to calculate it by hand. Probably can put that in the Appendix.
Validity of the findings: In the Result section, authors put a lot of texts describing the tables or the figures. These should be in the table/figure caption. For example: line 285-289, line 297-301.

Line 342, do authors think an improvement of 0.1 for R2 is a real increase?

The sample size is somehow small (60), although I understand a lot of work is needed in the field and lab for these 60 samples. An possible analysis could be performing a kriging based on the 60 samples to map out the whole area, and then you can pick out more than 60 points (say 100 or 200 points) with values calculated from the kriging in GIS, compared with data from UVA monitoring, so you will have more sample size.
Additional comments: This study is trying to examine if using unmanned aerial vehicles can help estimate soil salt content. It compares the soil salt contents estimated from unmanned aerial vehicles-borne multispectral imagery vs. the values from field sampling and laboratory analysis. It also compares three variable selection methods and different machine learning algorithms. Overall, I find this paper is accepted with revisions that I provided above. Mainly is increasing the sample size by performing a kriging to get more points of "true" values for the plot.


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: ESTIMATION OF SOIL SALT CONTENT BY COMBINING UAV-BORNE MULTISPECTRAL SENSOR AND MACHINE LEARNING ALGORITHMS
Review round: 1
Reviewer: 4

Output:
Basic reporting: The english is not clear and professional and does not meet the journal standards.

Literature and references do provide sufficient background.

Professional figures and tables are present

Self-contained with relevant results to hypotheses.
Experimental design: - Original Primary Research
- Research question is well defined
- Investigation does not sufficiently explain how spectral bands and indices of interest are related to the soil property in question. More fundamental explanation of the underlying chemistry is needed to justify research approach and demonstrate that the models aren't just fitting to noise.
- Additional details on the model parameters is needed in text. There sufficient explanation about features used, but not about the internal model parameters that were selected and why.
Validity of the findings: Train/test split is satisfactory and statistically sound.

Strong conclusions about the performance of the BPNN and SVR models are drawn in the results that are later tempered in the discussion. The strong conclusions in the results need to be removed. See detailed comments for more explanation.

The conclusions are relying on relatively arbitrary model performance criteria to justify success. The performance criteria need to be better justified.
Additional comments: Line 63: be more specific about what you mean by salt damage and remove “salt damage.
Line 100: specify what is meant by satisfying results
Lined 108-110: explain briefly to the reader how VIP and GRA are conducted
Line 168: This sentence is unnecessary
Line 176: Be specific about what treatments were run to remove impurities and what impurities were removed
Line 187: Good field of vision is subjective, be specific
Line 197: Explain why these bands were selected and what soil spectral properties they relate to in order to justify why they were selected.
Line 212: Define what DPS system is
Line 250: SVR is not really new
Line 261: Site the specific R packages that were used.
Line 266-267: These levels are commonly cited in the literature but there isn’t a scientific justification for why they are used. Also Nawar et al 2016 is not the originator of these criteria. Add a scientific justification for why RPD values were used
Line 274: 33, 40 and 27% don’t line up with none, mild and severe.
Line 300: Why was 0.7 selected
Line  323: You can’t conclude this from your study. You do not provide any detail about the BPNN parameters that were used. Neural networks are tough to fit, and you could just have an underfit model because of the specific model parameters used, not because of BPNN architecture generally.
Line 331: Define robustness
Line 352: Shortingcomings for this specific application. Again as with BPNN, SVR model turning can be very tricky and getting the model parameters right is difficult. Be careful in how much you generalize the performance you had to SVR models for this data generally. RF models are the easiest to tune which could be why you are seeing better results.
Line 366: Why were these bands better, what optical or chemical characteristics of the soil related to salinity are related to these features. Explanation is needed.


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: ESTIMATION OF SOIL SALT CONTENT BY COMBINING UAV-BORNE MULTISPECTRAL SENSOR AND MACHINE LEARNING ALGORITHMS
Review round: 2
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: This version has been well modified. The idea of using drones to monitor soil salinity is worth promoting. So I recommend to accept and publish.


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: ESTIMATION OF SOIL SALT CONTENT BY COMBINING UAV-BORNE MULTISPECTRAL SENSOR AND MACHINE LEARNING ALGORITHMS
Review round: 2
Reviewer: 2

Output:
Basic reporting: The authors did an excellent job of responding to my original comments. It is much better than before.
Experimental design: -
Validity of the findings: -
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: ESTIMATION OF SOIL SALT CONTENT BY COMBINING UAV-BORNE MULTISPECTRAL SENSOR AND MACHINE LEARNING ALGORITHMS
Review round: 2
Reviewer: 3

Output:
Basic reporting: None
Experimental design: None
Validity of the findings: None
Additional comments: This paper was well revised. I only have one more question. Line 362, judging which model is better based on R2 is not accurate. Usually R2 will increase with more variables being added into the model. Probably adding AIC although authors did add other two criteria (RMSE, RPD)


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: MEASURING RECOGNITION MEMORY IN ZEBRAFISH LARVAE: ISSUES AND LIMITATIONS
Review round: 1
Reviewer: 1

Output:
Basic reporting: The article provides sufficient introduction and literature references, however, I suggest some changes listed in the general comments below.
Experimental design: The experiments are well performed, however I have some concerns regarding the relevance of the study.
Validity of the findings: I suggest some changes in the Discussion and Conclusions sections. Please read the general comments for the author.
Additional comments: In this paper, Bruzzone et al analyze recognition memory in zebrafish larvae. The main conclusion of the study is that zebrafish larvae presents recognition memory, but some “non-cognitive” factors may complicate the assessment or interpretations of the tests. I found some difficulties understanding the relevance of the study and I have some concerns about the experimental design and the interpretation of the data.

While reading the abstract and introduction, I could not find a clear statement justifying the study. Authors list various papers from previous literature where recognition memory (NOR) has been found in zebrafish, but a clear explanation on the importance of assessing this memory in early stages of development must be provided.

In the introduction, I found the nomenclature regarding the developmental stages confusing. For the reader to have a quick and efficient comparison with previous reports, expressions as “third week of life” must be substituted by clear periods or days. To give consistency with the results section I would recommend changing everything to days post fertilization.

From line 53 to 61 in the introduction, authors listed a couple of papers where zebrafish are place-conditioned with an electroshock or pairing tail contractions with spotlights. The authors draw a justification for the present study by questioning the possibility that larvae can learn a “more advance” form of learning and memory. The claim that recognition memory could be considered a more advance form of learning must be logically justified and references for this must be provided. I suggest avoiding unclear concepts, especially on the justification of the study.

Please provide references for the statements on lines 69 to 72, on the advantages of using object recognition memory.

Experimental design and methods:
Please provide a clear justification for the selected post-fertilization days.

Please provide image resolution and sampling rates used in video recordings and provide a clear explanation on how these parameters are appropriate to track, with sufficient spatial and temporal resolution, the movement of the larvae.

Please provide a rational explanation to use two different novel object recognition tests (3D, Fig. 1A-B vs 2D Fig. 1D). In fact, I would be tempted not to consider the “Bi-dimensional” version as an object recognition test. This would be more similar to a place preference test, where different cues in the environment bias the preference of the subjects for one or other places. In this experimental design, subjects don’t have access to directly interact with the object. This conceptual clarification is necessary to avoid confusion in the interpretation of the data.

Some clarifications on the design of the experiment are needed. First, the dimensions of the box are different than the other experiments. This one is a smaller box; the reason of this change must be justified. Is it possible that as animals grow, they would feel less comfortable in a smaller environment?

Results

For all figures, asterisks indicating significant differences should be included.
In Experiment 1b, the preference index reported 55.88% + 21.98%, this is a very low index of preference, does that mean that not all the animals presented preference over the novel object? The use of parametric statistics and representations makes it even more difficult to understand. Is the data normally distributed? Please clarify and justify the use of parametric statics and representations for this and the rest of the data.
In line 293, the phrasing “marginally significant tendency” is not clear, in parenthesis the p = 0.073 indicates not significance.
In the meta-analysis (lines 297 to 309), please refer every subpanel of figure 5 in the text.

Discussion

I suggest addressing the possibility that the objects used in the study are not sufficiently different from each other.
Discussion must include a section illustrating the relevance of the main finding, that is, zebrafish larvae display higher levels of object recognition in the 14-dpf. How this data advances our knowledge regarding learning and memory? Please provide a clear statement on the pertinence of keep using this model in these developmental stages.
The findings of neophobia are in my opinion very interesting. Neophobia may represent a “complex cognitive” feature that may require, among other things, memory. For example, wouldn’t neophobia be a sort of object recognition test? How come an animal can avoid something new if it does not classify it as something new? I would suggest a few lines on the significance of this results and not only using it as a justification for the lack of object recognition after 21-dpf.
Conclusion must address the three objectives of the study (lines 85 to 87), on whether zebrafish larvae   display recognition memory, when this ability first appear in development and whether there’s a reliable method to asses it. Speculations on neurodegenerative disorders should be part of the discussion.


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: MEASURING RECOGNITION MEMORY IN ZEBRAFISH LARVAE: ISSUES AND LIMITATIONS
Review round: 1
Reviewer: 2

Output:
Basic reporting: See General Comments for the Authors.
Experimental design: See General Comments for the Authors.
Validity of the findings: See General Comments for the Authors.
Additional comments: The authors performed behavioral tests on zebrafish larvae to test if, and at which age, the larvae develop object recognition capabilities. Larvae of 7, 14 and 21 days post-fertilization were  tested on three variants of the Novel Object Recognition test (NORt). The results show that, overall larvae explored the novel object 53% percent of the time, a small but statistically significan effect. The results also the novel object recognition test peaks at age 14 dpf and decreases by 21 dpf, an effect that might be due to the behavior of neophobia developing at later stages.

Overall, the experiments and behavioral measurements seem well performed and the manuscript is adequately structured and written.

I have only one major question. Did authors rotate the location of the colors (and forms) so that, for example, blue could to the north but also south and west and east from the center of the petty dish/tank? This is important since preference for color of form might be confused by other environmental factors that made the larvae swim to a particular location of the petri dish or tank. Thus, is it important that the authors randomly change (or counterbalance) the spatial orientation of the behavioral apparatuses.

Minor comments:

-Line 26. Remove “that”.

-Line 96. The phrase needs rewording.

-Line 229. Define “IS”.

-In figure 3, please use lines and asterisks to illustrate the significant differences.

-Figure 5 show a preference for the green color, as opposed to the red preference mentioned in the text. Are labels wrong on the figure?


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: MEASURING RECOGNITION MEMORY IN ZEBRAFISH LARVAE: ISSUES AND LIMITATIONS
Review round: 2
Reviewer: 1

Output:
Basic reporting: No comment
Experimental design: No comment
Validity of the findings: No comment
Additional comments: The authors addressed all my previous concerns.


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: MEASURING RECOGNITION MEMORY IN ZEBRAFISH LARVAE: ISSUES AND LIMITATIONS
Review round: 2
Reviewer: 2

Output:
Basic reporting: No comment.
Experimental design: No comment.
Validity of the findings: No comment.
Additional comments: The authors addressed my comments and queries satisfactorily. Thank you.


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: DIRECT MEASUREMENT FOREST CARBON PROTOCOL: A COMMERCIAL SYSTEM-OF-SYSTEMS TO INCENTIVIZE FOREST RESTORATION AND MANAGEMENT
Review round: 1
Reviewer: 1

Output:
Basic reporting: The English, reviews and material is clear.  I question whether we really need the
lengthy recitation of patents.  This clogs the reference list and, to be honest, seems
difficult for me to understand what is being patented.  This is not my area!
Experimental design: Applying field eddy flux measurements to determining the effectiveness of a forest to sequester carbon is a new, clean idea. The methodology is oversold.  The limitations of the method are not as clearly presented as they should be.

I wrote my review as a piece, and have put this piece in section 3.
Validity of the findings: Review of:
“Direct Measurement of Forest Carbon Sequestration: A Commercial System-of-Systems to Incentivize Forest Restoration and Management”,
by Marino, Truong, Munger and Gyimah.

Reviewed by:  David Fitzjarrald, Atmospheric Sciences Research Center, SUNY Albany
			(N.B. I do not submit anonymous reviews.)

General comments.
My experience with direct measurements of carbon flux is that of a micrometeorologist. I’ve been around this work for some decades now. However, I have no background in the economics of financing carbon sequestration.  My main reaction in reading this material is that the first author has the inverse experience, and those co-authors who have the technical background have not fully argued the case for understanding a number of limitation to the method.

	Most of the issues that come to mind have to do with repeated claims that the eddy flux approach delivers values at half-hourly intervals.  That is the way the tables that the authors downloaded do present the data, but to assure that these numbers represent net ecosystem exchange, corrections must be made before the final data set is released. This is what rankles about the statement that the method is (line 247) “high precision, high frequency…” It will be more convincing when the authors outline how much better this approach is than competing ones, whatever they are.

These include: Correction for the chronic underreporting of nocturnal fluxes, which bias the results to excessive overall uptake, an error memorialized for all time by a paper by Grace et al. in 1990’s extolling exaggerated C uptake in the Amazon forest.  The correction in force for some years now has been simply to assert that the respiratory emission on windy, well-mixed nights holds for the other nights. Nearly twenty years ago we quantified that 10-20% of the CO2 escapes horizontally at Harvard Forest (Staebler and Fitzjarrald, 2004). There is another bias in filtering to fit into the half-hourly data window which can, on certain kind of days, underestimate daytime fluxes (Sakai et al., 2001). I am not certain how the error limits on the Harvard Forest data are estimated, but I would hope that these effects are included.  There is the ‘science’ of gap-filling—making up data when the instruments fail—introduces another problem.  Finally, the authors imply that the eddy flux method applies to entire enormous forests, but they are certain to know that the ‘footprint’ for flux measurements is only a small area—at Harvard Forest it has been reported to be 0.23 km2 (Kim, 2015 ). (The species composition at Harvard Forest For depends on wind directions, for example.)  Where is the discussion of the degree of homogeneity of the forest area being considered?  Much of the discussion of the Ghanaian forest has to do with the species richness and paucity of commercial timber.  These are truly important considerations but largely outside the purview of carbon uptake measurements.  Perhaps the authors do not take these issues to be ‘show-stoppers’ given their urgency to commercialize the product.  However, since critics of their commercial proposal could easily be as adept as am I, the authors should tighten their case.  As it stands, much of their text resembles what one might hear from a manufacturer of eddy flux equipment, Dr. Burba, for example.  Optimism is only sometimes a virtue.
	Another difficulty with this paper is the comparison of four years of data from the tower in Ghana with the longest eddy flux record anywhere, at Harvard Forest.  I believe that this paper would be greatly served by a careful stating of the uncertainties in the eddy flux method, a plan for assessing the similarity of the forest being considered with the ‘footprint’ of the eddy flux measurement being made. Most convincingly this would be done with some remotely sensed data.

	Please help me to see how the remuneration plan works on years, possibly long stretches of time during which a measured forest loses carbon over time. It appears that C uptake at Harvard Forest can be quite variable.  Last year, coauthor Munger et al. (2018) noted:
At Harvard Forest: “Annual NEE ranges from -0.4 to -6.1 Mg-C ha-1y-1 (always carbon uptake), with a mean of -2.9 (±1.5) Mg-C ha-1 y-1 .  Here we focus on the highest and lowest carbon uptake years, 2008 and 2010, respectively in order to examine how forest carbon balance can shift so dramatically over a 2 year interval.”
If the system were implemented, would the landowner have to pay someone back if the forest is a net carbon emitter?  What happens in the case of a wildfire?  (Actually, one could follow the other should there be a prolonged drought.)  Please try to address these issues.

Specific comments.

1. Line 141 “Genetic Heat Index” not common knowledge.  A reference is needed.
2. Line 181.  The absurd number of citations to patents fills up the bibliography.  Kindly explain to the naïve reader just what is being patented here. Put this information into Appendix 1.

References.

Kim J., 2015. Carbon and water cycles in mixed-forest catchments: ecohydrological modeling of the influence
of climate variability and invasive insect infestation, PhD diss. Boston University.

Munger, J.W., Whitby, T.G. and Wofsy, S.C., 2018, December. Rapid Shifts in Annual Carbon Balance For a Temperate Deciduous Forest: Validating and Diagnosing Ecosystem Surprises. In AGU Fall Meeting 2018 Abstracts.

Sakai, R.K., Fitzjarrald, D.R. and Moore, K.E., 2001. Importance of low-frequency contributions to eddy fluxes observed over rough surfaces. Journal of applied meteorology, 40(12), pp.2178-2192.

Staebler, R.M. and Fitzjarrald, D.R., 2004. Observing subcanopy CO2 advection. Agricultural and Forest Meteorology, 122(3-4), pp.139-156.
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: DIRECT MEASUREMENT FOREST CARBON PROTOCOL: A COMMERCIAL SYSTEM-OF-SYSTEMS TO INCENTIVIZE FOREST RESTORATION AND MANAGEMENT
Review round: 1
Reviewer: 2

Output:
Basic reporting: In general, the manuscript has gotten some potential to published in the PeerJ journal.
Nevertheless, some issues need to be considered before accepting the manuscript for publication. The manuscript needs some further improved before to be accepted for publication. In general, there are still some occasional grammar errors through the manuscript especially the article ''the'', ''a'' and ''an'' is missing in many places, please make a spellchecking in addition to these minor issues. The reviewer has listed some specific comments that might be helpful of the authors to enhance the quality of the manuscript further. Please consider the particular comments listed below!
Experimental design: •	The abstract is well written. The structure is fine.
•	Please remove the abbreviation.
•	The method section in general, is well written.
•	Please explain what was the data resolution used in this study (hourly, daily…) and why?
•	For better understanding of the methodology I would recommend adding a flowchart.
•	What type trees did the authors used to extract information about carbon concertation
Validity of the findings: •	The result section is well written.
•	Some sensitivity analysis is needed, perhaps using boxplots.
•	Figure 3, the maps are a low quality, please improve and scale bare. Legend and north arrow.
The discussion is a bit naïve, please elaborate it more. The discussion should provide a summary of the main finding(s) of the manuscript in the context of the broader scientific literature, as well as addressing any limitations of the study or results that conflict with other published work.
•	This section is well written, nevertheless is too short. Please include some quantitative findings and elaborate it further.
•	State the main findings resulted from this work through bullet points.
Additional comments: •	I would suggest to elaborate further this section by adding more literature.
•	 The objectives are not explicitly stated. It is not very clear what is the contributions of this work.
       Please explicitly state the main objectives of this work.


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: DIRECT MEASUREMENT FOREST CARBON PROTOCOL: A COMMERCIAL SYSTEM-OF-SYSTEMS TO INCENTIVIZE FOREST RESTORATION AND MANAGEMENT
Review round: 2
Reviewer: 1

Output:
Basic reporting: The revised version of the manuscript is greatly improved.
Experimental design: I appreciate that the authors now more clearly explain the difficulties in making the eddy flux measurement.  Their added text and citations makes their case a more viable, realistic-sounding approach.
Validity of the findings: I believe that the paper will not be improved significantly with much further revision.
Additional comments: See attached document for a few, additional but minor 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: DIRECT MEASUREMENT FOREST CARBON PROTOCOL: A COMMERCIAL SYSTEM-OF-SYSTEMS TO INCENTIVIZE FOREST RESTORATION AND MANAGEMENT
Review round: 2
Reviewer: 2

Output:
Basic reporting: Well done!
Experimental design: Well done!
Validity of the findings: Well done!
Additional comments: 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: URICASE-DEFICIENT RAT IS GENERATED WITH CRISPR/CAS9 TECHNIQUE
Review round: 1
Reviewer: 1

Output:
Basic reporting: In their article entitled "Uricase-deficient rat is generated with CRISPR/Cas9 technique" authors describe mechanisms of creation and main physiological parameters of uricase-deficient rat as a relevant model to study hyperuricemia in humans.  It is written in understandable manner, however, the language correction is strongly recommended. The manuscript has professional structure with sufficient background; most recent relevant literature is cited; figures and tables illustrate the obtained results. Raw data are shared. Manuscript describes very valuable research model important for the scientific community. The conclusions are supported by the obtained data. Although the manuscript cannot be recommended for publication in the present form, I would recommend the revision.
Experimental design: The aim of presented study fully meets Aims and Scope of the journal. The absence of relevant models of hyperuricemia taken together with growing prevalence of this disease form one of the biggest problems in modern physiology. The present research describes an elegant model relevant for investigating hyperuricemia in humans. All procedures described in the study are performed according to the existing ethical guidelines and approved with local ethical committee; modern molecular techniques and adequate statistical approach are appropriate. Some methods (Uricase activity assay, Western blot, RNA integrity assessment) should be described more detailed.
Validity of the findings: No comment
Additional comments: Design and generation of model
Lines 112-113: are both RNA sequences match forward strand of gene as it is stated?
Line 128: Please, specify the mentioned molecular techniques.

Blood and organ sample:
133: please, specify the volume of blood sample
138: you should specify the calculation of left kidney index here (in the place of first mentioning)
140-141: What do you mean "any surviving rats"? After described procedures all rats should be euthanized.
Uricase activity assay - please, describe more thoroughly.
Western blot - please, describe the procedure and protein standards used more thoroughly.
Line 154 - Please, specify, which protease inhibitors cocktail has been used. Roche makes several. Also add data on manufacturers.
Line 170 - RNA integrity assay should be described more thoroughly.
SUA, BUN, Cr, blood fat and serum glucose determination - please, do not use SOP abbreviation for instructions(protocols) which are provided by manufacturers.
Histological examination of kidney sections
Please, specify the microscope. Please, provide histological scores for changes observed, as in present form data from histology do not seem to be considerable, as "sporadic changes" could be observed even in healthy animals.

Statistical analysis:
Have you analyzed the data distribution?
Lines 210-211. Please, do not comment on difference if P = 0.456. There is no difference.
Please, comment on birth and mortality rate of offspring. Please, specify the time when they were born.
Lines 232-235: Please, transfer to the Materials and Methods section.
Lines 241-242, 246-249: Transfer to Discussion.

Histological examination:
There are ways to assess tissue status numerically. Please, make it, otherwise the presented results do not support your conclusions about differences in kidney structure observed in uricase-deficient rats.

Conclusions:

I would like to avoid such strong comments as "an ideal model"


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: URICASE-DEFICIENT RAT IS GENERATED WITH CRISPR/CAS9 TECHNIQUE
Review round: 1
Reviewer: 2

Output:
Basic reporting: Field background: the description of the functional role of the gene deleted in the present study, the pathway in which it is involved, and the evolutionary differences between animal species is inaccurate and lacks relevant literature citations.
Experimental design: The inclusion of data on urate renal excretion would have completed the mutant characterization.
Validity of the findings: The authors explanation for the observed differences in Uox-deficient mice and rats is not convincing.
Additional comments: The manuscript by Yum, Zhang et al. reports on the generation and analysis of a Uox-deficient rat. The rat model is apparently healthy although with elevated serum urate. This is an important result as the Uox-deficient mouse, the current model for hyperuricemia, is a suffering animal. The rat model could also be advantageous in experimental protocols benefiting from a larger animal. The characterization of the rat mutant  is well performed, but it would have been more complete and informative with the inclusion of data on urate renal excretion. Overall, the experiments are well described (but see below) and the availability of raw data adds value to the work. Specific points are listed below in the order they appear in the manuscript.

Line 3. “Nanzhang”. Correct?

Page 7. Introduction: “Uric acid can be further transformed by uricase to allantoin…”. This is an old notion that does not account for current knowledge of the urate degradation pathway. For reference, see the Metacyc entry: https://biocyc.org/META/new-image?object=PWY-5691.
Uox is a pseudogene in apes, not  in all primates. Recently it has been reported that Uox is inactivated also in elephants and manatees (https://doi.org/10.1093/nargab/lqz012). Contrary to what is stated by the manuscript, Uox is not a pseudogene in reptiles and birds, although its function in uricotelic vertebrates is unknown.

Pag. 10 “Enough uric acid was dissolved in 50 mM ...”. Please be more specific and provide at least a quantity range.

Line 184. Please provide identifiers for the kit used in the determination of SUA, BUN, Cr, etc and the name of the manufacturers.

Pag. 12. “Because of an associated with an off-target site” (?). Rephrase.

Pag. 13. “SUA was elevated (Fig 4B)”. Is the increase significant? Please rephrase as “SUA was elevated compared to wild type rats (see Fig. 4B)” and assess the significance of the comparison.

Pag. 13. “less than 20 μg/ml (Table 1), and not higher than 40 μg/ml 219 (male) or 30 μg/ml (female)”. If SUA is less than 20 μg/ml, it is not surprising that it is not higher than 40 or 30 μg/ml. Also, if the authors did not observe significant differences for male and female SUA in Uox-/- or Uox+/-, why did they use different cut-offs for male and female?

Pag. 14, line 245. This chapter confoundingly entitled “mRNA splicing” is unclear. My guess is that here the authors have aligned short reads obtained by next-gen sequencing to the rat Uox mRNA sequence using Blast and the TBtools package.  It is not clear which particular procedure included in the TBtools package they have used. It is also unclear how “reads were spliced together to puzzle out the full length of Uox mRNA”. The term “splicing” here is improper. Please refer to the vast literature on RNA-seq mapping (e.g. doi.org/10.1186/s13059-016-0881-8) for the specific programs and descriptions for type of analysis. Please use “nt” instead of “b” as an abbreviation for nucleotides.

Line 283. Can the authors provide a reference for “normal ranges” in the rat serum?

Line 302. If the smaller dimension of mouse is an explanation for the higher SUA in the Uox-KO mouse compared to the Uox-KO rat, how can the mouse SUA be comparable to that of humans? A more likely explanation for the observed SUA differences are differences in the renal excretion/reabsorption of urate.

Line 306. Please correct “humane males”.

Fig. 1. Use absorbance at 292 nm (Abs292) instead of optical density (OD). At time 0, the Abs (OD) is reported to be 0, while serum and liver homogenate are expected to have a non negligible absorbance. Is the quantity on the Y-axis a delta Abs?
According to the authors, there are “similar patterns” of uricase activity in liver and serum. This is strange as the Uox activity should be present only in the liver. Consistently, decrease of absorbance in Fig. 1A (blue line) starts immediately, while in Fig. 1B starts after 100 min and is probably not due to enzymatic degradation.

Fig. 2. Both Western Blot (A) and RT-PCR (B) lack controls with housekeeping genes. It would have been interesting to see the same analysis with the Uox+/- rats.

Fig. 3. The Figure does not represent “mRNA splicing”. It shows the alignment of RNA-seq reads to the rat Uox mRNA sequence. The meaning of the striped bars should be clarified.

Fig. 4. Comparison with age-matched wild-type rats is lacking. Please correct “USA” in panel B.

Table 2. Please report the number of offspring that survived for each breading.


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: URICASE-DEFICIENT RAT IS GENERATED WITH CRISPR/CAS9 TECHNIQUE
Review round: 2
Reviewer: 1

Output:
Basic reporting: The authors have satisfactory addressed most of the requests. The description of field background has been improved, methods have been specified, and additional figures and tables have been added to the results.
Experimental design: Although the authors did not present data on urate renal excretion, this issue is now discussed in the present version.
Validity of the findings: The authors have refined their hypothesis about the differences in urate serum level in different mammals. They maintain, however, an explanation based on the  "faster cell cycle" of small mammals. I think that speaking about higher "metabolic rate" and  "cellular turnover" of small mammals could be clearer for the reader.
Additional comments: Although the authors state that the meaning of striped bars in Fig 3 was clarified, I could not find this explanation in the revised version. Graphics in Figure 3 are different from those presented in Raw data (in which striped bars are not present).


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: YEARLING PROPORTION CORRELATES WITH HABITAT STRUCTURE IN A BOREAL FOREST LANDBIRD COMMUNITY
Review round: 1
Reviewer: 1

Output:
Basic reporting: na
Experimental design: na
Validity of the findings: na
Additional comments: This manuscript point out a potential interesting development to correlate yearling proportion from constant-effort landbird capture stations with habitat structures and which potentially can be used as a proxy for habitat quality. The manuscript is well written but I have some critical suggestion for improvement. Overall the manuscript lack ecological knowledge, this is necessary to be able to understand the species specific responses. Additionally a more thorough deviation of species into groups regarding their habitat preferences or functional groups is needed. This was done in figure 4 but this is unclear and should also be included in the methods section.

The use of proportion and probability combined is confusing throughout the manuscript. In my mind probability tells you the likelihood of something happening (model predictions), while proportion is just the comparison of (measurable) quantities, thus they do not tell the same and should not be mixed. In line, the title and research question are linked to yearling proportion but your models and figure 4 representing probabilities (predictions from the model). This results in the fact that you relate the probability that you can catch a yearling with habitat structure and not the proportion. Please clarify this throughout the manuscript. Find below some general comments:

Comments:

Line. 74. What about including sex? Is there a potential effect of sex on yearling proportion, this may differ between species, but competition and habitat exclusion may act different on male and females.

Line. 75. A little bit more information regarding species is required. Potential correlations between yearling proportions and habitat structure depend on habitat requirements of the studied species. More general ecology should be included and additionally it may be useful to define functional groups or groups representing their habitat requirements.

Line 79-80. I my opinion the second goal cannot be achieved within the current study setup. Habitat quality was not measured by a more reliable/known proxy and thus you cannot examine the accuracy and utility of yearling proportion as a measure of habitat quality. Just habitat (EVI, Greenness) not enough to classify high or low quality habitat, more precise measurements of for example reproduction is needed.

Line 98. Operating the stations until 7 August seems quite long. I am not completely familiar with the breeding season in this area but in general in July the nestlings fledged and birds start to move around more. This mean that your bias in capturing local breeding birds increase (higher influx from non-breeders and possible individuals from other areas after nest failure). Do you for example get the same result if you only use the data from June? Or can you correct for this effect in the models?

Line 174. Linear regression assume normal distribution of the data, even for time series. Thus in this case, could you provide extra information regarding this, I expect that data with N=5 cannot be normally distributed. If so, linear regression cannot be used, you can consider to use non-parametric correlation (Pearson correlation) instead.

Line 185-187. Why do you calculate probabilities as your research question is about proportion? I see you report probabilities in the result section L. 222-223, but you discus proportions in the discussion L. 260-263.

Line 222-230. Grouping the species in habitat classes is excellent but this is not written anywhere else in the intro or method. It would be good to know which species belong to which group and how they ecologically are divided.

Line. 238-259. Arrange according to the research question in the end of the introduction.

Line 248. Although habitat quality is a widely used concept, it can be misleading and vague. I think that within this study habitat quality is poorly defined and used in the wrong context. Within this study 29 species are used, all with differences in niche occupation. I think it is to straight forwards to say that EVI can track habitat quality, this is in my opinion species specific. Therefore this should be formulated in the way as the title indicate “Yearling proportion correlates with habitat structure” which is not similar as habitat quality. However, I do agree that yearling proportion can be an indication of habitat quality but this need more support.

Page 24, Figure 4. Be clearer about the species groups.


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: YEARLING PROPORTION CORRELATES WITH HABITAT STRUCTURE IN A BOREAL FOREST LANDBIRD COMMUNITY
Review round: 1
Reviewer: 2

Output:
Basic reporting: The paper “Yearling proportion correlates with habitat structure in a Boreal forest landbird community” reports bird banding data, specifically the proportion of yearlings, and its relationship to habitat factors in landscapes ranging from natural boreal forest to reclaimed land of different ages.
The paper is very well written and interesting. As far as I can judge, the statistical analyses are sound. I have only a few comments requiring some clarification:
L137: What was the spatial scale on which habitat was classified in situ?
L161: What was the rationale behind using exactly this spatial scale? Have the authors also evaluated other spatial scales or can they give reasons/references why this scale was considered?
L214: It is not entirely clear what is meant by "showed little development". Please rephrase and clarify
L238: It is unclear to which finding this first sentence is referring to. If this is just a general staement, consider removing it. I guess there are even some birds breeding  in land which has not been reclaimed yet? Some ground breeding species? So, this statement makes - probably - only sense when referring to forest species
Experimental design: no comment
Validity of the findings: no comment
Additional comments: No additional comments


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

Input:
Title: YEARLING PROPORTION CORRELATES WITH HABITAT STRUCTURE IN A BOREAL FOREST LANDBIRD COMMUNITY
Review round: 2
Reviewer: 1

Output:
Basic reporting: na
Experimental design: na
Validity of the findings: na
Additional comments: Dear authors,

Many thanks for adjusting the manuscript according to the suggested improvements. All the points of concern are adjusted to satisfaction and thus the manuscript is ready for publication.

There is one small detail: Reference in line 58 (Saracco, DeSante, & Kaschube, 2009) should be checked! In line 58 year of publication is changed to 2009, while in reference list (line 418-419) indicate 2008.

Kind Regards,


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: YEARLING PROPORTION CORRELATES WITH HABITAT STRUCTURE IN A BOREAL FOREST LANDBIRD COMMUNITY
Review round: 2
Reviewer: 2

Output:
Basic reporting: My previous commets were only minor and have been all well addressed
Experimental design: no comment
Validity of the findings: no comment
Additional comments: No additional comments


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

Input:
Title: IDENTIFICATION OF EXOSOMAL MIRNAS ASSOCIATED WITH THE ANTHRACYCLINE-INDUCED LIVER INJURY IN POSTOPERATIVE BREAST CANCER PATIENTS BY SMALL RNA SEQUENCING
Review round: 1
Reviewer: 1

Output:
Basic reporting: No comment
Experimental design: No comment
Validity of the findings: Authors reported interesting findings on exosomal miRNAs associated with the
anthracycline-induced liver injury. Seven subjects were recruited and showed significant differences in ALT. To find the potential AILI-related exosomal miRNAs, authors tried to integrate their findings with known DILI-related genes. I have several concerns about this study as following.
Major
1.	The study only screened the circulating exosomal miRNAs. No further functional experiments were conducted. So, the authors should state the type of tissues used in the CTD Database and explain the potential regulatory mechanisms.
2.	In the method part, authors should demonstrate their criteria or threshold in each step. For examples,
a.	in “ALT detection and grouping of patients”, they should state the threshold for grouping and the reasons or references for choosing this threshold;
b.	in “MicroRNA sequencing and data analysis”, what is the threshold for purity of RNA?
c.	In data analysis part, are the p-values raw values or adjusted values? If adjusted values, which method is used to adjust the p-values?
d.	How many exosomal miRNAs were totally identified using small RNA seq?
3.	The gene enrichment analysis looks unnecessary for this study. Because the reference dataset is DILI-related genes. All the terms should be just related to DILI.
4.	In discussion part, authors only used around five sentences at the end of this part to discuss the major findings in their study. They may consider searching if the previous studies find same miRNAs related to AILI, if the same target genes were found, if the regulatory type (up/down-regulated) was same.
Minor
1.	Authors should carefully check the paper again. Like in line 276, “has-miR-503-5p” should be hsa-miR-503-5p.
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: IDENTIFICATION OF EXOSOMAL MIRNAS ASSOCIATED WITH THE ANTHRACYCLINE-INDUCED LIVER INJURY IN POSTOPERATIVE BREAST CANCER PATIENTS BY SMALL RNA SEQUENCING
Review round: 1
Reviewer: 2

Output:
Basic reporting: 1. Many abbreviations were not explained in the manuscript. Such as APAP (line 286), NAFLD (line312), etc.
2. In Figure 3C, the number of non-overlapped miRNAs should also be presented.
Experimental design: 1. The major concern is that the limited number of patients in the study. Only 7 were recruited: 2 were classified as non-injury and the other 5 were classified as injury. It will significantly lead to cases of bias. More patients need to be recruited to get a more solid conclusion. This limitation should also be discussed in the section of discussion.
2. No experimental evidence were included in the manuscript. Although the author stated that the aim of the work is to identify potential miRNAs, the absence of experimental results makes the work incomplete and less convincing.
Validity of the findings: 1. The author should clarify the number of replicates for each sample when performing RNA seq (at least two). The other information should also be included such as the quality of the sequencing and the read depth and length.
2. Line 184, the author should explain the meaning of inference score, and justify the reason to use 100 as the cutoff, considering that this an import step in the analysis.
Additional comments: The manuscript of Zhang et al. has identified differently expressed exosomal miRNA from 7 post chemotherapy patients. They performed small RNA seq and bioinformatics analysis. They suggested that miR-1-3p and miR-503-5p are potential targets for further study. Although the language is clear, several major flaws make it not suitable to be accepted for publication.


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

Input:
Title: IDENTIFICATION OF EXOSOMAL MIRNAS ASSOCIATED WITH THE ANTHRACYCLINE-INDUCED LIVER INJURY IN POSTOPERATIVE BREAST CANCER PATIENTS BY SMALL RNA SEQUENCING
Review round: 2
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: Generally, authors have answered my questions thoroughly. I think that the design of this study is clear and the finding is meaningful.


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: IDENTIFICATION OF EXOSOMAL MIRNAS ASSOCIATED WITH THE ANTHRACYCLINE-INDUCED LIVER INJURY IN POSTOPERATIVE BREAST CANCER PATIENTS BY SMALL RNA SEQUENCING
Review round: 2
Reviewer: 2

Output:
Basic reporting: Now the manuscript is in a good form to be published.
Experimental design: The issues have been addressed.
Validity of the findings: The findings are substantially improved.
Additional comments: Manuscript of "Identification of exosomal miRNAs associated with the anthracycline-induced liver injury in postoperative breast cancer patients by small RNA sequencing" has been improved substantially over an earlier 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: A SCOPING REVIEW OF DETERMINANTS OF PERFORMANCE IN DRESSAGE
Review round: 1
Reviewer: 1

Output:
Basic reporting: Generally a very thoughtful review that has high relevance for equine researchers and practical use for the field of equestrian dressage in practice. The literature has been well selected for the purpose of the review, which was to identify quantitative measurements for dressage performance for both the horse and/or the rider. This review was very well written, and the text flowed logically.

To further improve the reading of the manuscript, I would recommend to start the first sentences of the introduction with a reference to para dressage or dressage, similarly to the abstract, as the first paragraph of the introduction takes quite long to reach the topic of the review, (Para) Dressage.
Experimental design: This was a well-designed systematic literature review.

I am missing the aspect of equine welfare, and rider skills in relation to welfare. Specific riding methods, such as Rollkur, which are mentioned in your keywords, and their effect on kinematic variables and/or welfare of the horse fail to appear in the discussion. It could be argued that horses whose welfare is impacted have potentially lower performance. Was there a particular reason why this aspect did not appear more in the discussion?
Validity of the findings: Based on my knowledge, all reported findings and conclusions are valid. I would like to see in the limitation section a discussion of the limitation of performance scores as a reference for performance, given the sometimes low agreement between judge scores (e.g. Stachurska and Bartyzel, 2011).

I have a few specific questions about statements:

Line 238-241: This is very interesting! However, I am missing the link between energetic efficiency and performance. It would also likely be more efficient for some horses to transition to a faster gait instead of lengthening the stride, but adapting stride length and frequency irrespective of energetic efficiency is a main requirement for dressage. Could you please expand on the aspects of efficiency, regularity and the vectors of braking and propulsion found to be relevant to dressage by Biau and Barrey (2004)?

Line 267-269: Is the step frequency at the walk necessarily rapid? Does it not also depend on the ability of the horse and rider to increase stride length and decrease stride frequency?

Line 302: what do you mean with “tracking length”? do you mean “over-tracking distance”? It would be nicer to have consistency in the terms used, especially as you use “over-tracking” in the following sentence.

Line 303: “longer strides are achieved by increasing propulsion and the propulsion vector…” Is the propulsion coming from the hind limb? In what direction is the propulsion vector going? Please further explain this.

Line 314-315: “these traits are regarded positively by judges”. Please provide references for this statement or combine the sentence with the previously cited results from Biau and Barrey (2004).

Line 324: Why and how? This statement should be supported by references.

Line 340: “creating impulsion”. Do you mean “impulsion” in the equestrian sense or in the biomechanical sense? It could be helpful to define this more thoroughly, maybe in lines 290-292, where the term first appears.
Additional comments: I have some further comments on the manuscript:

Specific comments:
Abstract:
Line 38: is there a difference between “posture” and “postural position”? “Postural position” seems to me like a word redundance. Also on line 357.

Introduction:
Line 50: it would be helpful to start the first sentence with a reference to Para Dressage, and then go on to explain how Paralympic disciplines are defined. The paragraph takes quite long to reach the topic of the review, (Para) Dressage. It is much nicer in the beginning of the abstract (line 14).
Line 76: for walk, you reference only Clayton 1995. Back et al. 1996 also analysed the walk in relation to gait quality. This reference may be added.
Lines 90-103: this is where the welfare aspect of rider skills could be discussed.

Survey methodology:
Line 155: included instead of include.
Line 156: selected instead of included?
Line 159, 160: Remove space between the backslash and the second word.
Line 160: what is “adjustability”? This term has not been previously described and is probably not common. Please define the term.
Line 160: same comment for “connection”. Please define the term.
Line 174: please remove space between the backslash and the second word.

Discussion:
Line 219: “to improve their performance” instead of “to develop an improved performance”.
Line 219-222: This statement needs one or several references.
Line 348: “Lusitanos” instead of “Lusitano’s”
Line 347-249: These breed specific differences are interesting. It would be nice to add a few references on breed-specific differences in locomotion (e.g. Barrey et al. 2002).
Line 357: “postural position” like a word redundance. Would “posture” not be enough?
Line 443-452: very nice!

Table S1: the expression is "throughness", from the German “Durchlässigkeit”, and not "thoroughness".


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

Input:
Title: A SCOPING REVIEW OF DETERMINANTS OF PERFORMANCE IN DRESSAGE
Review round: 1
Reviewer: 2

Output:
Basic reporting: See below
Experimental design: See below
Validity of the findings: See below
Additional comments: General comments
The manuscript presents a review of scientific evidence related to dressage performance. The text is well written, with very few typographical errors.
The aim of providing a scientific background for classification of disability levels in para-dressage is legit, but this goal is little touched upon in the discussion. I started my reading with the search strategy, and read through to the end of discussion before I returned to the introduction, and I was rather surprised when I read the incentive for the study. I also wondered whether this review of the literature was asked for by the Paralympic committee or if this was an initiative by the authors themselves.
The review adds moderately to the existing body of literature. Similar material is available in book chapters and previous reviews mentioned in the current manuscript, though all of this information is not available in the same paper, as the authors point out.

The study is reported as a systematic review, but I would argue that what has been undertaken here is not a systematic review, rather it’s a scoping review. A systematic review summarizes multiple studies that have addressed the same research question, while a scoping review maps the existing literature relevant for a particular question or area of interest. It is obvious already from the aims, that this study addresses a broader area rather than a single focused question and the results section does not present evidence to inform practice, rather it’s presents mapping of the literature available and overview of study findings. This is not suggesting a lower value of the publication, but I feel that it’s important to present it in the correct framework, and this review also does not benefit from trying to squeeze itself into the systematic review framework where it clearly doesn’t fit. The guidelines of this journal refers to the following publication for guidance whether a review is a systematic review or a scoping review.

 Munn Z, Peters MDJ, Stern C, Tufanaru C, McArthur A, Aromataris E. 2018. Systematic review or scoping review? Guidance for authors when choosing between a systematic or scoping review approach. BMC Medical Research Methodology 18:143 DOI: 10.1186/s12874-018-0611-x.

I suggest that you read this publication. Related to this issue, use of the PICO formula does not fit a scoping review, this search strategy is mainly tailored towards systematic review of intervention studies and in your form you state intervention was non-applicable in your case.

Materials and methods
The initial search strategy appears non-optimal, the chosen combination of words could pose a risk that relevant papers are missed. However, I interpret that you extended the search using keywords that appeared relevant when you went through your search results. Is that a correct interpretation? It is difficult to follow the search development from the supplement. I am not sure what the numbers listed actually mean, it seems it’s the number of hits for each word. I think some explanatory notes would be good to make this more interpretable. Also, why is the VetMed sheet empty?
I miss the prisma checklist, which is mandatory for systematic reviews according to the journal guidelines. But as I understand this is not mandatory for scoping review.
Your inclusion criteria seem to suggest that every paper needs to contain something on both horse and rider. Is that really the case? These criteria are also not easy to understand. I suggest that you include an example after each except the first one. For number two you can for example write e.g. optic motion capture/ kinematic data.
It would be good if you state when the search was performed, even though this information is given implicitly by the time span of the search.

Results
Please add journal, volume and page numbers to your list of included papers if feasible. This will make the supplements more valuable resources for readers.

For me, figure 2 is difficult to read, there is a lot of arrows crossing. Are the relationships illustrated in figure 2 based on findings in the reviewed papers or suggestions by authors of these papers, and/or your own reasoning? Please be clear about this in the text and figure legend. Regardless, I think these interrelationships are open questions, apart from the few that we have data on. For example, in the text you say that ’stiller leg will improve the rider’s ability to provide consistent and precise aids to the horse, resulting in more finite speed, gait and/or movement changes’ in trot while the figure suggests that less knee flexion-extension does not have an influence on joint kinematics.

Discussion:
I feel the discussion could be more condensed and be more concluding and interpreting rather than just reporting. There is no clear connection or reference to figure 2. I don’t mean that you have the rewrite the whole discussion, but I suggest that you go over the text with this in mind.

Line 245: please add that the comparison between a smaller and a taller horse is only applicable if they are walking at the same speed. Unless I misunderstood this in someway.

Line 267: please add a reference for rider pelvis pitch rotation.

Line 268: I’m not sure if I agree with this. Both walk and trot have two cycles of rider pitch per stride, and in trot the stride frequency is higher. Further, depending on the speed of the respective gaits, walk step frequency is not that much higher compared to trot.

The conclusions are comprehensive and well written, and seem relevant. But the discussion lacks connection to the conclusions. If you go through the discussion according to my suggestion above, I hope this will improve.

Supplement
One experimental study is classified as before and after. According to my understanding before and after studies are non-experimental. If you have a different understanding please provide (me) a reference.


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

Input:
Title: A SCOPING REVIEW OF DETERMINANTS OF PERFORMANCE IN DRESSAGE
Review round: 2
Reviewer: 1

Output:
Basic reporting: As before, the clarity of language was excellent.
Experimental design: The authors have adapted their description of the review (scoping instead of systematic) according to reviewer two's comments. The comments I had personally on the aspect of equine welfare were addressed in the discussion/conclusion.
Validity of the findings: All my comments were addressed in the revised edition of this manuscript. Thank you.
Additional comments: I just have two small comments:

on line 322: there is a space missing between "horses" and the reference.

on line 370: the reference is not complete "(Barrey et al.,"