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: LEPTIN ADMINISTRATION DOES NOT INFLUENCE MIGRATORY BEHAVIOUR IN WHITE-THROATED SPARROWS (ZONOTRICHIA ALBICOLLIS)
Review round: 1
Reviewer: 2

Output:
Basic reporting: The authors investigated effect of leptin hormone administration in migratory bird (white-throated sparrow; Zonotrichia albicollis) under long day photoperiod (14 hours of light and 10 hours’ dark) during pre-migratory. The objective was to test if leptin administration can induce fat deposition and early appearance of night time migratory restlessness.  The manuscript has a few caveats that need to addressed or resolved with caution. Authors have interpreted results and drawn conclusions based on low sample size and limited time points that has been acknowledged in discussion. It is an interesting study but need major revision to improve the presentation and clarity in different sections of the manuscript.
Experimental design: Author acknowledges here that studies on avian leptin has been controversial. I think all the studies in past including this work done in this manuscript uses murine leptin to test its response on avian physiology withought testing how similar murine and avian leptins are in terms of chemical nature, protein sequence, and other biochemical properties.
Are there any reports showing cross reactivity between antibody raised against murine leptin and avian plasma isolate?
My biggest concern is what if avian and mammalian leptin are structurally very different. Knowing it is very different, what is the rationale behind testing its role in avian physiology?

The experimental design is OK and author has mentioned the caveats in discussion.
Validity of the findings: The objective was to test if leptin administration can induce fat deposition and early appearance of night time migratory restlessness.
As expressed earlier, my biggest concern is what if avian and mammalian leptin are structurally very different. Knowing it is very different, what is the rationale behind testing its role in avian physiology?
Experimental design, and statistics used are fine.
Additional comments: Line 42: you can add Singh et al., 2018, Frontiers in physiology showing fat deposition in liver in migratory birds a relevant reference to show how fat is deposited in migratory state and it disappears post-migration.

Line 51: However, leptin ………. balance hormone, has been……migration. The first and half sentence is not connected. Reframe the sentence.

Line 58,59: Break the sentence into two. The message is not clear from the sentence. I guess author wants to mention here that avian leptin could not be sequence earlier due to high GC content and that also makes it challenging to amplify with standard PCR.
The end of the sentence and low expression is not clear here in the sentence.

Line 64: Structural modelling……. Hydrophobic core. What information author want to give here. It doesn’t connect with the previous sentence.

Methods & results-

Line 118: We captured 24 white-throated sparrows. Author used morph in running statistical model and results. It is not clear throughout the method what was the number of different morphs in the analysis.
Also, give power estimates for statistical significance.

Another question: did author corrected individual mass for body size. I think there could be individual body size differences which was not taken into account for body fat.

Line 168: correct recordings spelling in the sentence.

Discussion

Line 227: Hormones like ………..influence migratory activity. I think author should include Mishra et al., 2017 Hormones & Behavior showing role of circulating corticosterone and insulin levels in night time activity migratory (daily levels and rhythm in circulating corticosterone and insulin are altered with photostimulated seasonal states in night-migratory blackheaded buntings).

Adding some information in discussion on structural, biochemical differences on mammalian and avian leptin would help in conjunction with how difference in the structure could be a potential cause of no functional relevance of leptin in birds.


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: LEPTIN ADMINISTRATION DOES NOT INFLUENCE MIGRATORY BEHAVIOUR IN WHITE-THROATED SPARROWS (ZONOTRICHIA ALBICOLLIS)
Review round: 2
Reviewer: 1

Output:
Basic reporting: The article has much improved from the first draft. I have two more requests to further improve the clarity of the article:

1) Can you please comment in the discussion on whether you've managed to induce a migratory state at all? The lack of change in fat and the DECREASE over time in activity suggest that you may not have. That would change the interpretation a little bit, as part of the interpretation is that birds stop responding to exogenous leptin when in migratory state. Are there other studies that have used photoperiod to induce migratory restlessness in this species and do they see a different pattern?

2) You gave some really nice and clear explanations in the response to reviewers that could also be added (without much effort) to the manuscript itself:
- The explanation of how models are averaged
- The reassurance that after transformation the data met the assumptions of the models

Given the changes made, maybe the title and abstract should talk about "Leptin treatment" rather than just "Leptin"?
Experimental design: This has been properly dealt with.
Validity of the findings: I have no problem with the data or the interpretation, with one exception: the difference in activity between the two morphs is interpreted as a difference in migratory restlessness. Given the doubt that any migratory restlessness was induced and there was no interaction between trial and morph, maybe it's safer to interpret that as difference in activity levels only.

I do also wonder why for the fat data, a "normal" two-way (repeated measures) ANOVA was used, and no the same model selection approach as for the behavioural outcome measures. Sex and morph could affect fat as well, surely? But if there is a good reason, then there is no need to change this.

Finally, in the responses to reviewers, the method of validating the EthoVision data was explained. Please explain that here too. And, if I understand the validation method correctly, can a rank correlation not show this validity?
Additional comments: A few small points:
- why are the tables numbered 1.1, 1.2 and 1.3? Just number them 1, 2 and 3.
- when saying there is a significant effect of morph on activity levels, please tell us which is more active
- line 176 of the Word (tracked) version: "recordings" instead of "recordins"
- line 193 of the Word (tracked) version: please remove "used"


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: ABOVE- AND BELOW-GROUND TRAIT COORDINATION IN TREE SEEDLINGS DEPEND ON THE MOST LIMITING RESOURCE: A TEST COMPARING A WET AND A DRY TROPICAL FOREST IN MEXICO
Review round: 1
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: Data analysis should be improved.
Validity of the findings: In this manuscript, the authors compared the aboveground and belowground traits of a dry forest and a wet forest, and want to find if the traits differences are formed by the environmental stress. The trait differences between two forests are listed clearly in the manuscript, but they are not correlated with the environmental factors analysis. The authors cannot make conclusion about if the traits differences are necessarily formed by the environmental factors or which factor simply. The analysis also did not show which environmental factor is the most limiting resource influenced the species traits. My suggestion is that the correlation analysis between environmental factors and traits are needed instead of simple traits differences comparison. To improve the manuscript, the theoretical frame about how environmental factors limiting traits variation should be also strengthen.
Additional comments: The manuscript could be improved by strengthen the theoretical frame and data analysis.


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: ABOVE- AND BELOW-GROUND TRAIT COORDINATION IN TREE SEEDLINGS DEPEND ON THE MOST LIMITING RESOURCE: A TEST COMPARING A WET AND A DRY TROPICAL FOREST IN MEXICO
Review round: 1
Reviewer: 2

Output:
Basic reporting: In this paper the authors analyze the importance of functional traits in seedlings of 94 tropical tree species in two contracting ecosystems. It is an interesting and current topic for understanding plant responses to changes in the environment. The introduction documents the topic adequately; however, throughout the paper the citations used are not very recent and more than five years old, there are only 3 citations from 2020-2021. So I recommend updating the introduction and discussion, some articles that could support are the following.

Stem functional traits, not just morphology, explain differentiation along the liana–tree continuum
LS Santiago - Tree Physiology, 2021
El Niño-Southern Oscillation affects the water relations of tree species in the Yucatan Peninsula, Mexico J Palomo-Kumul, M Valdez-Hernández, GA Islebe… - Scientific Reports, 2021
Hydraulic traits of Neotropical canopy liana and tree species across a broad range of wood density: implications for predicting drought mortality with models ME De Guzman, A Acosta-Rangel, K Winter… - Tree Physiology, 2021
Unraveling the relative role of light and water competition between lianas and trees in tropical forests: A vegetation model análisis F Meunier, H Verbeeck, B Cowdery, SA Schnitzer… - Journal of Ecology, 2021
Experimental design: no comment
Validity of the findings: no comment
Additional comments: I recommend replacing table 2 with table S1, since I consider it very important to have the list of species studied in the paper. Table 2 should be placed in the supplementary material.
I consider it necessary to add in Table S1, which traits were measured for each species? Since, in the methodology they mention that only root data were obtained for 28 species in the dry forest.


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: ABOVE- AND BELOW-GROUND TRAIT COORDINATION IN TREE SEEDLINGS DEPEND ON THE MOST LIMITING RESOURCE: A TEST COMPARING A WET AND A DRY TROPICAL FOREST IN MEXICO
Review round: 1
Reviewer: 3

Output:
Basic reporting: Figures must be supplied at a greater resolution.
Experimental design: The description of how traits were measured is completely missing. I understand that this detailed description is in a supplemental material, but a brief description should be included in the main text. Furthermore, statistical analyses can be summarized in the main text, and explain in more detail in the supplemental materials.
Validity of the findings: This work seeks to understand trait coordination and trade-offs among plant organs in species from two tropical forests: one dry and one moist. The authors studied a large number of species and traits related to plant performance and found coordination in both sites, but the traits involved in those coordinations differed between sites. This is an important contribution to understanding plant function in tropical forests, demonstrating that trait coordination in the dry site involved traits related to water acquisition and storage, as water is the main limiting resource in those forests.

Data have not been provided
Additional comments: Major comments
The introduction has enough background information to understand the motivation behind this work. I suggest in the PDF that the authors check other papers on the coordination of traits among organs. Also, given that coordination and trade-offs are frequently used throughout the manuscript I recommend that the authors define these terms at first mention in the introduction.
The description of how traits were measured is completely missing. I understand that this detailed description is in a supplemental material, but a brief description should be included in the main text. Furthermore, statistical analyses can be summarized in the main text, and explain in more detail in the supplemental materials.
Results are concise and well describe. I added a comment in the discussion related to the relationship between SLA and SRL. Please revise that the growth-defense trade-off is appropriate for explaining these results.
Minor comments
I have included minor comments directly in the PDF attached.
Please supply images of better resolution.
Please include data availability statement and mention if the data is published elsewhere or if it will be published as supplemental materials.
I also recommend that the authors include pictures of the plants/sites in the supplemental materials.


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: HARMFUL EFFECTS OF THE MICROPLASTIC POLLUTION ON ANIMAL HEALTH: A LITERATURE REVIEW
Review round: 1
Reviewer: 1

Output:
Basic reporting: The english language and writing quality is unequal along the paper. The introduction especially needs particular attention:
L51 "The" term plastic
L57 1mm thick
L82 "different classes of animals" or "differentanimals"
L93:  terms «soil», «air», «water», or «ocean» were used in the query
L114 microparticles of any polymer type
L140 I suggest  “Microplastics Environmental pollution”
L323 define ICR at first occurrence

Literature is sometimes missing or not cited early enough.
L165 References for the content of plastics in River?
L176 reference of the article in sweden?
L180 reference for the MP in the air?
L211 refernce for the MP amount per year
L213, is this a percentage you calculated among all studies ? Or did you fin in a review?
L239 Give the name of the review in the first sentence of the paragraph

The structure is globally fine. Some minor change could be made:
L72. I recommend you first introduce the plastic composition with main polymers and additives and then the properties.
L.201 I would present the MP in food and finish with the human feces and placenta

The review is of broad and cross-disciplinary interest and within the scope of the journal

 The field been reviewed recently. I suggest the authors focus on the plastic content in animals and the potential effects

Additional figures and discussion would increase the interest for the review
L260, add a table and/or figure to describe the effects in fish

L331 discuss the applied doses and related effects in comparison with the environmental content in the matrices (soil, water) and animals you describe previously

L337 It would be nice to have a figure to summarize the effects on mice

Please also discuss other factors leading to toxicity that are not yet tested, for example the transport of sorbed contaminants or pathogens on the plastic particles and the degradation stage of the plastic.

The title should be much more specific as the subject developed by the authors is clearly the effect of microplastics in animals.
Experimental design: The Methods are not described with sufficient detail & information.
L96 How many original papers and reviews were published on these topics?
 L118 Why studies until 2017 are selected if you say you analysed from January 1st, 2010 onwards?
Validity of the findings: L65. Why would you include “particles formed as the result of automobile tires wear, some types of road surface” as primary MP. I doubt small plastic particles are deliberately created in these case.

L72. I suggest you also mention physical properties such as elasticity and shear strength. These properties will depend on the plastic polymers so I recommend you first introduce the plastic composition with main polymers and additives and then the properties.

L133 for water, dissolved sea salt and honey I would be surprise a digestion step would have been included. Pease check and give references.

L137 I don’t think you mean “Then” may be “alternatively”: spectral method (FTIR or Raman) and visual counting give both the number of MP

L182 I would not consider cellulose fibers as MP
Additional comments: I thank the authors for their review of this very interesting topic. The authors describe the potential for microplastic in animals and expected harmful consequences. I suggest to define better the scope of the review, in the title and introduction.


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: HARMFUL EFFECTS OF THE MICROPLASTIC POLLUTION ON ANIMAL HEALTH: A LITERATURE REVIEW
Review round: 1
Reviewer: 2

Output:
Basic reporting: In general, the manuscript reviewed few aspects: methods for determining the content of microplastics, microplastics in water, soil and air, microplastics in food chains and human food, hazardous effects of microplastics on the health of different classes animals (Invertebrates, Fishes, Mammals). It has a broad interest, but it didn’t make a solid foundation to estimate the potential risk of microplastics to human health.

The title is too general. As the authors stated in the manuscript, “The aim of this review is to systematize and summarize the results of experimental studies of the microplastics effect on the health of different classes animals and to estimate the potential risk of microplastics to human health.” Why don’t you focus on your aim and only review the hazardous effects of microplastics on the health of different classes animals?
Experimental design: no comment
Validity of the findings: The argument for the goals set out in the Introduction is not well developed. More statements are needed for how to link the results from animals to human.
Additional comments: Line 24, undoubt to undoubted. Line 60, hundreds to hundreds of. Line 95, full stop dot (.) is missing in the end of the sentence.
Please check the number in Line 153 and 157, are they 268.94 and 79.000 or 26894 and 79000?
Lines 185-193, please add references where the data from.
Line 231, please rephrase the subtitle “Microplastic hazard effects on animals”. It’s not correct in gramma.
Line 243, please rephrase “in the presented in the review works”.


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: HARMFUL EFFECTS OF THE MICROPLASTIC POLLUTION ON ANIMAL HEALTH: A LITERATURE REVIEW
Review round: 1
Reviewer: 3

Output:
Basic reporting: The article is conformed to professional standards of courtesy and expression. It is written in clear, unambiguous English. Nonetheless, at line 114 “tested microparticles of any plastics” could be written in a clearer way as well as “Microparticles of plastic” in line 198, that could be simply referred as “Microplastics” or “MP”.

The article includes sufficient field background with appropriate references providing justification for this literature review. As a minor review, please check reference 62 (line 605): a full stop after “2021” is missing.

Improving the description of nanoplastics may be beyond the purpose of this manuscript, nonetheless I suggest adding NP abbreviation at line 61 when “nanoplastics” are first mention so that it is possible to recall them explicitly through the text when it could be used (lines 263, 264, 266, 328 and in Table 1).

The structure of the article is conformed to an acceptable format of ‘standard sections’ of the Journal. However, the title could be more specific, stating that the review is focused on MP effects on animal health with special attention on mammals or laboratory rodents.

The abstract fulfils its scope, as a minor review, please specify that the “Small particles of plastic” (line 17), are of different types.

The Table 1 is relevant to the content of the article summarising the results from different studies, however it should be improved to make it easier to consult and more exhaustive:

- All the “Ref #” numbers have to be changed to be properly associated with the actual references.
- The “Animals“ column should be compiled in a consistent way through the Table. The colour code “Red text – female animals” needs to be checked in Luo T et al.,2019(a) and (b), Rafiee M et al.,2018, Park EJ et al., 2020 and Lim D et al., 2021. “Male” has to be added in Lu L et al., 2018 and Jin Y et al., 2019.
- The heading of the column “Type and size of MP particles” should be stated in a more suitable way since in some cases different treatment and NP are included. When NP are used it should be made explicit (Qiao J et al., 2021, Rafiee M et al., 2018, Fournier SB et al., 2020 and Estrela FN et al., 2021).
- Please use English alphabet in the “Dose” column in Xie X et al., 2020.

The review is of broad and cross-disciplinary interest, as it is also stated by the authors in lines 27-28. Although the field has been reviewed recently enough, the value of this article, that describes the field and explains why it is needed, stays in the different point of view that provides. As a matter of facts, it considers the results from the most recent papers about the study of MP effects in vivo, on laboratory rodents, which can give new insights on effects on human health as it is made clear since the introduction (lines 83-87).
Experimental design: The Survey methodology is well described. It is reproducible, the used criteria ensure comprehensive and unbiased coverage of the field and the assessment of the updated state of the art. The sources are appropriately cited and the manuscript is logically organized into subsections. As minor revisions:

- Details about the MP entrance mechanism in the women placenta (Ragusa et al., 2021) (lines 203-204) can be added.
- In lines 321-322 can be mentioned that different types of MP and NP are also taken in account in Table 1.
- In line 328 can be indicated that plastics particle with a diameter of 0.02 are considered NP.
Validity of the findings: The conclusions are appropriately stated and connected to the original question investigated. Nonetheless they can be expanded. It can be discussed if the doses tested in the in vivo studies are comparable to the MP potential exposure in the natural environment and/or giving an indication for approaches and future directions for further research.
Additional comments: No additional comments


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

Input:
Title: HARMFUL EFFECTS OF THE MICROPLASTIC POLLUTION ON ANIMAL HEALTH: A LITERATURE REVIEW
Review round: 2
Reviewer: 1

Output:
Basic reporting: Despite some improvements, some part of the text are still difficult to read because of the grammar. I listed some errors below but please ask a fluent speaker to carefully review all the document.

L16 “Small particles of different plastic types”
L17. Connect the two sentences : “Small particles of different types plastic with a diameter of less than 5 mm are found around the globe in water, air, soil, and various living organisms. An individual constantly absorbs microplastics with water and food, inhales with air.

L21. I think this sentences are un-necessary in the abstract: “Experimental studies of the potential health risks of microplastic are now under the undoubted scientific interest. The number of publications on this topic is growing every year. However, it could be difficult to analyze the potential data because of narrow investigations and contradictory results obtained, which should be combined and generalized.

l26 "of different classes animals"

Figure 1. Brain “behavior in shallow water” is not connected to the rest of the sentence.

Figure 2 connect the sentence “MP accumulates in liver and kidneys causing inflammatory and oxidative stress” or remove verbs “MP accumulation in liver and kidneys, inflammatory and oxidative stress” and harmonized in all the figure.

Figure 2 I suggest to highlight the specific organs in the text with bold, as it is done in the Figure 1

l. 403. “Plastic waste broken into small particles of MP, which carried by water and wind” verbs are missing
l404: “particles of microplastics with food” reformulate. Suggestion “Water, food, and inhaled air are pathways for microplastics to penetrate and accumulate in the bodies of various living organisms, including human.”

L407: “can provide”  -> “can have”
Experimental design: The citation has been improved.
Validity of the findings: Table 1 and Figure 1 and 2 are good tools to systematize and summarize the microplastics effect on the health
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: HARMFUL EFFECTS OF THE MICROPLASTIC POLLUTION ON ANIMAL HEALTH: A LITERATURE REVIEW
Review round: 2
Reviewer: 2

Output:
Basic reporting: The article is conformed to professional standards of courtesy and expression. It is written in a clear, unambiguous English.
Could you please ensure that “MP” is written in a consistent way through the text? A couple of times it is written “MPs” and it is not clear to me if it is done just by mistake or as a purpose.

The structure of the article is conformed to an acceptable format of ‘standard sections’ of the Journal. The title is specific, stating that the review is focused on MP effects on animal health.
The abstract fulfils its scope.
The Table is relevant to the content of the article summarising the results from different studies.
The Figures describe appropriately the MP effects on animals.
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: ANTIDMPPRED: A WEB SERVICE FOR IDENTIFYING ANTI-DIABETIC PEPTIDES
Review round: 1
Reviewer: 1

Output:
Basic reporting: The introduction section provided a good background to antidiabetic peptide therapeutics, setting the scene for the application of machine learning approaches and the objectives of the study. A little bit more background on BioDADPep would have been useful, particularly as this resource was used as the exclusive source of antidiabetic peptides in the data collection.

However, the paper is lacking a view of comparable tools or services. The authors state that there are no other anti-diabetes peptide predictors, and that is true, but it should be possible to appraise wider developments that are relevant e.g. taking a glance across at anticancer peptide predictors to see what kind of methods they use and what kind of benchmarking scores they're putting out. It's not a like for like comparison but would yield valuable reflection and context.

This is particularly important as the abstract states "surpassing other state-of-the-art classifiers" but there is no evidence presented for that claim, indeed as made clear in the introduction "there is currently no classifier for predicting such peptides". There is perhaps a contradiction here, unless what the authors mean by "other state-of-the-art" is their own SVMs and kNN that they implemented in the paper (which performed less well than Random Forest), which in that case as currently written is misleading and should be clarified.

Figure 1 should be associated with the methods section rather than the introduction.

The cited papers were well referenced and relevant. Ensure consistency with no use or use of hyphen in "antidiabetic".

Good use of figures and the legends are clear. I'm not sure about the visual resolution. The tables are fine.

Raw data fine.
Experimental design: The study represents original primary research within the scope of the journal. The important research question was very well defined, as is the targeted knowledge gap. The methods were outlined clearly.

Methods - with the AVPdb database, how was it ensured that the "control" non-antidiabetic peptides had no antidiabetic activity in reality? If this detail is contained in the annotation of AVPdb, then this should be explained. If, however, information from the AVPdb can not be used to confirm whether peptides are in reality devoid of any antidiabetic property, this is an aspect that should be considered in the design of the study. Otherwise, the study could be inherently incorporating false negatives inappropriately in the training sets. "Negative" sets are sometimes really "unknown" sets and whether this is the case should be clarified here - and either way, how well that maps to the desired use of the tool and its interpretation.
Validity of the findings: The findings appear valid, though the question relating to ensuring that the "control" non-antidiabetic peptides had no antidiabetic activity in reality needs to be addressed.

The study resulted in the development of a useful tool. The reported accuracy of 77.12% and area under the receiver operating curve (AUCROC) of 0.8193 in the nested five-fold cross-validation is good, but by no means exceptional, for ML classifiers and might be further improved. Data augmentation techniques, such as those used in this paper - ACP-DA: Improving the Prediction of Anticancer Peptides Using Data Augmentation https://www.frontiersin.org/articles/10.3389/fgene.2021.698477/full - might be used to improve the scores somewhat.

Though there is no reason to doubt the stated statistics for performance for the "known antidiabetes vs known not" sets, when we tested a CYP2D6 sequence chopped up into 30-aa "peptides", the following predictions were generated (please see uploaded image) - 13 peptides were predicted to be antidiabetic peptides, 4 not. This is a very high level of positive prediction for what is "nonsense" data in this regard.

This is probably due to the Random Forest classifier not having been trained on data like this, and so it is generating a high proportion of "low confidence" predictions (e.g. near 0.5), marginal "yes" decisions, though some scores are markedly higher.

I note that the website suggests applying a modified threshold of "0.95" to "discover novel peptides with greater confidence", with a box allowing manual adjustment of the threshold. This is also mentioned in passing in the paper itself and appears to be a very important feature to increase the confidence we can have about the positive predictions.

This brings us to the central point regarding stated accuracy - is it possible to produce analysis and discussion about how the model's various accuracy statistics change, as the threshold changes? e.g. What's the accuracy, sensitivity, and precision of the approach when a user chooses 0.90, 0.95, 0.99? This analysis should be accompanied by a discussion of the envisioned use-cases for a 0.5 threshold vs. a 0.95 threshold, for example.

This would be valuable as it would allow users to know what level of greater (real-world) confidence in positive predictions they can actually have when following the recommendation to use the system at e.g. "0.95" vs the default "0.5".

This would give the user a way to attain some degree of certainty (e.g. t=0.95 might mean lots of false negatives but a very high PPV, so if you ever get a "yes" you can be sure it's a strong "yes" and is worth investigating further). This area needs further elaboration.

The AntiDMPpred web service itself is a tidy web development that appears to work effectively. It's fast with a nice straightforward interface.

There were no papers cited in the results and discussion section. The findings should be linked back to the relevant literature, which as alluded to above, needs to be expanded upon.

The conclusions are generally straightforward and measured. However, there is no way to corroborate the statement "AntiDMPred......will play a critical role in improving the development of useful therapeutic antidiabetic agents". It could indeed play a pivotal role because it is the first machine learning that has successfully targeted this research question, which is commendable enough in itself.
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: ANTIDMPPRED: A WEB SERVICE FOR IDENTIFYING ANTI-DIABETIC PEPTIDES
Review round: 1
Reviewer: 2

Output:
Basic reporting: The manuscript, AntiDMPpred: a web service for identifying anti-diabetic peptides, by X. Chen, B. He, and J. Huang, reports a new web-based tool that predicts anti-diabetic peptides. This tool has the potential to aid researchers in identifying lead peptides for further studies of bioactive molecules aimed at the treatment of diabetes.

The manuscript is assembled in a professional manner, with sections appropriate for the content of the study. It is self-contained, and the amount of content is appropriate for one publication unit. It is organized, generally well-written, and conforms to professional standards of the scientific literature. Below I have made a few specific suggestions about wording, grammar, and clarity.

Line 62  Avoid the word “defective” because that’s not really the case. How about: “Since the current therapeutic medications have significant drawbacks...”

Line 64  “Many peptides have been demonstrated to have impressive antidiabetic effects.”

Line 69  Unclear what this means: “scattered in bioinformatics resources.”

Lines 70-71  “Fortunately, BioDADPep, a database dedicated to storage and management of antidiabetic peptides, have been proposed.” What does it mean that the database is proposed? Do you mean it has been developed? Can you say one more sentence about what it is and what it does?

Line 95  “...where 2025 peptides had between 5-50 amino acids.”

Line 127  “For a peptide sequence with L amino acids, if k = 0,...”  (Small “i” in “if”)

Line 138  Unclear what this means: “There could be a part of key features which make a significant contribution....” Are you trying to say that the features needed to be cleaned up into order to ixdentify any significant contributions to the prediction or anti-diabetic peptides?

Line 143  “which were generated”

Lines 207-209  I suggest: “The RF/Pearson combination outperformed the other five combinations for all evaluation indicators except AUCROC.”

Lines 215-217  I suggest: “It is observed that the RF-based predictor outperformed the other three other machine learning predictors for all seven evaluation metrics. All seven evaluation metrics reached the highest level.”
Experimental design: Here I must insert the disclaimer that I am not a computational researcher, and I do not know a lot about computational or bioinformatics methodology. I hope that the other reviewer(s) was/were able to critically analyze the methods used in the study. For my part, I can say that the experimental design was clearly described and seemed to follow the conventions of the particular practices employed (at least based on the things I looked up while I was reading). It also appeared to me that the experiments were described in enough detail that they could be reproduced by another researcher who has a knowledge of computational methods. Additionally, the investigation appeared to be performed with high technical and ethical standards. In terms of identifying the knowledge gap that is filled by this contribution, the authors clearly stated that AntiDMPpred is the first tool capable of identifying antidiabetic peptides from sequence information.

Here are some suggestions to improve the reader’s understanding of the methods used to construct the datasets and extract the features:

Lines 88 + 94  Can you provide a little more information/description about both the BioDADPep and AVPdb databases?

Line 89  Can you say more about non-natural peptides versus natural peptides? How did they all come to be in the BioDADPep database? Why did you exclude the non-natural ones?

Line 131  Where does the number 25 come from?

Lines 132-133  This is too short to be a paragraph. Can you expand it with a little more description of the feature vector methodology? For example, where does the number 519 come from?
Validity of the findings: Again, I rely on the other reviewer(s) to evaluate the validity the computational findings, but I believe they were presented very clearly. I just have one question about the description of the results: in lines 218-219, why couldn’t the ROC curve be drawn or the AUCROC be calculated?

I did practice using the online tool, and I thought it was convenient and easy to use. I would suggest making the “Predict” button stand out more; perhaps it could be bright green. With all the buttons being gray, I felt I was naturally drawn to click on the “Browse” button simply because it is higher up, even though I was not uploading a file. I think a bright color that stands out for the “Predict” button would be very helpful.
Additional comments: This paper is a well-constructed and smoothly written, and by all accounts AntiDMPpred will be very useful web-tool. With the small corrections above, I think it will turn out to be an excellent 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: ANTIDMPPRED: A WEB SERVICE FOR IDENTIFYING ANTI-DIABETIC PEPTIDES
Review round: 2
Reviewer: 1

Output:
Basic reporting: The points that I raised in the previous review have been resolved by the authors.
Experimental design: The points that I raised in the previous review have been resolved by the authors.
Validity of the findings: The points that I raised in the previous review have been resolved by the authors.
Additional comments: I thank the authors for their constructive, thorough and meaningful rebuttal and for their care in the revisions that have been made to the manuscript. The research has proceeded from an innovative idea to the implementation of a very useful resource, now reported in an excellent paper.


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

Input:
Title: ANTIDMPPRED: A WEB SERVICE FOR IDENTIFYING ANTI-DIABETIC PEPTIDES
Review round: 2
Reviewer: 2

Output:
Basic reporting: The manuscript, AntiDMPpred: a web service for identifying anti-diabetic peptides, by X. Chen, B. He, and J. Huang, reports a new web-based tool that predicts anti-diabetic peptides. This tool has the potential to aid researchers in identifying lead peptides for further studies of bioactive molecules aimed at the treatment of diabetes.

The manuscript is assembled in a professional manner, with sections appropriate for the content of the study. It is self-contained, and the amount of content is appropriate for one publication unit. It is organized, generally well-written, and conforms to professional standards of the scientific literature.

I thank the authors for the corrections they made based on my first review, as well as that of the other reviewer. For the newly added text, I only have two very small suggestions about wording:

Line 331-331: Remove the word “receiving”: “There has been growing interest in using machine learning...”

Line 335: Add the word “an”: “However, an anti-diabetic predictor is currently lacking...”
Experimental design: Here I repeat the disclaimer that I am not a computational researcher, and I do not know a lot about computational or bioinformatics methodology. Clearly, the other reviewer was able to critically analyze the methods used in the study and make helpful suggestions that were addressed by the authors. From what I can surmise, the experimental design is clearly described and seemed to follow the conventions of the particular practices employed (at least based on the things I looked up while I was reading). It also appears me that the experiments are described in enough detail that they could be reproduced by another researcher who has a knowledge of computational methods. Additionally, the investigation appears to be performed with high technical and ethical standards. In terms of identifying the knowledge gap that is filled by this contribution, the authors have clearly stated that AntiDMPpred is the first tool capable of identifying antidiabetic peptides from sequence information.

The authors have addressed all the suggestions and concerns I put in my review. If the other reviewer is satisfied with how they addressed his suggestions, then I am also content.
Validity of the findings: Again, I rely on the other reviewer to evaluate the improvements the authors have made based on his review.

I thank the authors for adjusting the “Predict” button. I think the interface is appealing and easy to use.
Additional comments: My first review was generally positive with only small suggestions, and the authors have successfully addressed all my concerns thoroughly. Very nice work!

Here I have only included two very small wording suggestions, and I would not need to see the article again to be confident in its acceptance.


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 COMPARISON OF NON-SURGICAL METHODS FOR SEXING YOUNG GOPHER TORTOISES (GOPHERUS POLYPHEMUS)
Review round: 1
Reviewer: 1

Output:
Basic reporting: Overall fantastic, my only recommended additions would be a citation for adding lidocaine to the rostal et al 1994 methodology (Hernandez-Divers SJ, Stahl SJ, Farrell R. An endoscopic method for identifying sex of hatchling Chinese box turtles and comparison of general versus local anesthesia for coelioscopy. JAVMA. 2009;234(6): 800–804; Rakotonanahary TF, Kuchling G, Routh A. In-country anaesthesia for endoscopic sexing of the ploughshare tortoise (Astrochelysyniphora). Solitaire. 2015;26:1-14; emmel et al; Evan S. Emmel, Samuel Rivera, Freddy Cabrera, Stephen Blake, and Sharon L. Deem "FIELD ANESTHESIA AND GONADAL MORPHOLOGY OF IMMATURE WESTERN SANTA CRUZ TORTOISES (CHELONOIDIS PORTERI)," Journal of Zoo and Wildlife Medicine 51(4), 848-855, (12 January 2021).

I really like the component of the conclusion mentioning practicality of endoscopic sex identification in field/less controlled settings, and you could again reference Rakotonanahary et al 2015 and emmel et al 2021 in this section if references are to be included in conclusions.
Experimental design: No comment
Validity of the findings: No comment
Additional comments: Excellent work, very well-written study with an exciting contribution to the field of chelonian captive breeding and sex identification.


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 COMPARISON OF NON-SURGICAL METHODS FOR SEXING YOUNG GOPHER TORTOISES (GOPHERUS POLYPHEMUS)
Review round: 1
Reviewer: 2

Output:
Basic reporting: The language used throughout the ms is clear and concise and does an excellent job summarizing the literature and the gap that the study attempts to fill.
While it is unfortunate that there is not an easier way to sex neonates, the extraordinarily high rate of success for hatchlings is very encouraging and should be very useful for researchers working on a variety of GT management practices.

I made some small comments about the tables/figures in the PDF.
Experimental design: Excellent job in the methods providing detail and clarity for a series of complicated protocols.
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: A COMPARISON OF NON-SURGICAL METHODS FOR SEXING YOUNG GOPHER TORTOISES (GOPHERUS POLYPHEMUS)
Review round: 1
Reviewer: 3

Output:
Basic reporting: The writing is clear and unambiguous. The manuscript is well written. I found a few editorial items that I detail below.

Introduction and background are well referenced and relevant.
Figures are excellent, in high quality and clearly present the data. They are well labelled and described.

Raw data were supplied and I examined them.

On line 263 there is a discontinuity in the sentence

On line 313 the present tense is used when it should be past tense.

One line 468 the first word should be capitalized
Experimental design: The research is within the scope of the journal.
The research questions are well defined, relevant and meaningful.
The complicated design is well explained.
The research is performed to the highest technical and ethical standards
The methods are very well described and there is sufficient detail for the experiments to be replicated.
Validity of the findings: This is a beautiful study! The need for it is clearly stated in the Introduction. While the laparoscopic method is the gold standard for sex determination of freshwater turtles and tortoises, there is a need (well documented in this manuscript) for less invasive methods.This study addresses this need and clearly tests several different approaches. In addition, the methods are tested with tortoises of different ages so that makes the study even more valuable. The results are clear and the findings are of fundamental importance for both the biology and conservation of tortoises. The results are applicable to other species as well.

The underlying data are provided, are robust and statistically sound.

The conclusions are well stated and are linked to the original research question.
Additional comments: I wish I had thought of and done this study. It is a masterful job and a real tribute to both innovative science and science geared to the problems of conservation and climate change.

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: AGGRESSIVE SIGNALING AMONG COMPETING SPECIES OF BIRDS
Review round: 1
Reviewer: 1

Output:
Basic reporting: The manuscript is well written. Authors provide a good picture of the aggressive signaling literature; the only reference that I think should have been cited is Caro & Allen (2017, https://doi.org/10.1098/rstb.2016.0344). The manuscript is well structured although it feels a bit convoluted at some points because various analyses were performed without it being clear how each of them relates to the overarching question “what signals do birds use in aggressive contests among species and how do these signals vary across diverse groups of birds?” (particularly the within-species and within-family analyses). Raw data are provided with appropriate sources to back them up. I believe there is an excess of figures, some of which (e.g., Fig. 1, at least some panels in Fig. 2, Fig. 9) could be appended as Supporting Information.
Experimental design: The authors did a great job in identifying and making a case for how interspecific aggressive signaling constitutes a knowledge gap within the field of animal communication. Methods are described with sufficient detail and codes ran smoothly after I corrected a typo (signaling spelled as “signalling” in several instances, causing R to not identify the correct column in the data set). I have numerous concerns about the study design (ranging from data collection through data scoring to data analysis), which I detail in the General comments below.
Validity of the findings: In my view, data are not statistically sound because no analysis accounts for phylogenetic non-independence (see below). I did not inspect conclusions in detail as I have concerns about the methods that generated the findings that should support them in the first place.
Additional comments: In this manuscript, the authors aimed to provide an overview of the signals that birds use during agonistic interactions with heterospecific individuals. To that end, the authors analyzed photos and videos of mostly Neotropical species in aggressive contexts and described several aspects of their gestural displays.

The authors make a good case for why we should give more focus to interspecific aggression signals. The costs of fighting with a heterospecific can be just as dramatic as with a conspecific, so, just like for intraspecific signals, we should expect selection for interspecific signals that can resolve conflicts without physical combats. At the same time, there is reason to expect that interspecific communication should be under distinct selective pressures because of the differences in morphology, perception, and cognition of potential receivers. Since interspecific signals are understudied in comparison with intraspecific signals (a point also made by Caro & Allen 2017, https://doi.org/10.1098/rstb.2016.0344, which the authors should consider citing if they wish to make their case stronger), I believe the authors identified a relevant knowledge gap.

I also commend the authors on coming up with a nice system for classifying the different aspects of gestural displays across species, which ultimately contributes to a very interesting data set. The authors also provide detailed descriptions and illustrated examples for each category in the system, which is of great help to understand how data were scored.

Having said that, I have a number of concerns mostly about how the study was designed, which I will detail below.

I. Statistical approach
I invite the authors to reflect on the following question: why did they decide to use statistical tests?

Statistics are a set of tools that allows us to infer conclusions about a population from a sample of that population. Perhaps the authors are instead interested in making qualitative descriptions pertaining to their sampled species alone, without extrapolating the estimated parameters to the whole population (i.e., all bird species). I say that because the authors do not need a statistical test to be able to state, for example, that “the majority of birds in our dataset had a forward-facing position” (L219). This is a demonstrable fact: in the majority of their sampled interactions, the signaling bird had a forward-facing position. The authors could qualitatively describe the patterns they found for their sampled species (Figures 5–7 seem like a step in that direction) and use them to speculate on how birds in general behave during interspecific aggressive interactions and/or to suggest future directions for the field. That is what non-systematic literature reviews often do.

However, the fact that the authors chose to use statistical analyses tells me that they are interested in extrapolating their findings to birds in general. The problem is that, when we are comparing species, we necessarily have to account for the fact that some species are expected to be more similar to each other because they shared much of their evolutionary histories as a single lineage. In other words, the more recently two lineages diverged from the same branch, the more similar they should be simply because there was not enough time for them to become too different. This makes species non-independent data points that contradict the assumption of independency of traditional statistical tests like the ones used in the manuscript. The problems arising from not incorporating phylogenetic non-independence have been vastly demonstrated since at least Felsenstein (1985, https://doi.org/10.1086/284325) and I am afraid that the authors would necessarily have to account for it if they wish to extrapolate their findings to birds. A nice and thorough overview of phylogenetic comparative methods that do account for phylogenetic non-independence can be found in Garamszegi (2014, https://www.springer.com/gp/book/9783662435496).

If the authors indeed intend to extrapolate their findings to birds in general (with caution, given their small sample size), I suggest that they use phylogenetic generalized linear mixed models (PGLMMs) for all of their analyses. PGLMMs are nothing more than GLMMs that restrict species-specific intercepts to be more similar to each other according to how related species are in the phylogeny. I am not completely familiar with how PGLMMs are implemented in frequentist packages like lme4 but suspect it should be fairly straightforward. A quick search tells me that packages pez and phyr apparently include functions that do so, although I have never used them. Alternatively, function phyloglm in the package phylolm implements phylogenetic logistic regressions, but I am not sure how flexible it is in regard to the other random effects. In lme4-like syntax, the formula would be as follows:

response ~ predictor + (1|interaction id) + (1|species) + (1|phylogeny),

where “response” would be binary (is position assumed? is region/color highlighted? is region closest to receiver? is signal same/similar/different?), “predictor” would be categorical (postures, body regions, colors, signal similarity degree), “interaction id” and “species” would account for repeated measurements from the same interaction and species, respectively (both needed because some interactions involve the same signaling species), and “phylogeny” is the correlation matrix deriving from the phylogenetic tree. The authors can readily sample full bird trees from a Bayesian set available at birdtree.org.

II. Data search
The authors did an excellent job in providing the source for each of the analyzed interactions, but we are given no information on how they searched for data in the first place. Questions that I would like to see answered include: what media/article databases were searched and when? What searching strategies did the authors use? Did they use sets of keywords or did they focus searches on selected species, thereby prioritizing taxonomic diversity (L96)? More importantly, what precise criteria were used to determine whether a picture/video depicted an aggressive display? This last question is important because circular reasoning could be a problem if the criteria are not independent of the scored traits. For example, if open-wings posture, but not closed-wings, was a criterion to consider a behavior as an aggressive display, open wings would surely be well represented in the sample simply because they were used as a criterion, not because they are indeed common as aggressive displays.

III. Color analysis
I have two concerns regarding the color analysis. The first is again related to circular reasoning. In L127, one of the criteria used by the authors to classify a region as highlighted was one “that appeared to highlight contrasting patterns or coloration (e.g., black/white contrasting patches, black throat, bright red crest)”. In other words, the color of a region could determine whether the region was highlighted or not. Later on, naïve observers scored the color of those regions, meaning that colors were used both as a criterion for what constitutes a highlighted region and as a variable that describes that region. If authors automatically considered red, black, and black/white regions as highlighted, they were already biasing the results of the subsequent analysis, which unsurprisingly showed carotenoids, black, and black/white patterns as among the most highlighted colors. My suggestion is that the authors use only the first criterion for what constitutes a highlighted region (“those that were most obviously displayed or modified for display to the receiver”, L125) because it is independent of color. Otherwise, I am afraid that I do not see how the results from this analysis could have much use, unfortunately.

The second issue with this analysis is that colors were scored as 0 (“not highlighted”) even if they were not present at all in a species. An individual can only highlight colors it possesses, which is a sufficient — and biologically uninteresting — explanation for why, for example, albatrosses never highlighted carotenoids or structural colors. It also makes the analysis extremely sensible to the species that ended up in the final sample. For example, one explanation for the fact that rufous/chestnut is rarely highlighted is, say, that birds choose not to highlight this color because it is not recognized as a universal aggressive signal. But an equally likely — and again biologically uninteresting — explanation is that most species in the sample simply did not have any rufous/chestnut in the plumage and therefore could not possibly highlight it. There is no way to distinguish between these two explanations with the way highlighted colors were scored. My suggestion to make this analysis biologically relevant is to score colors as follows: (1) color is present in the plumage and is highlighted; (0) color is present in the plumage and is not highlighted; (NA) color is not present in the plumage. NAs would of course be dropped from the GLMM analysis. By doing as I suggest, 0 will have the unambiguous meaning of “not highlighted” (rather than “either not highlighted or not present”) and thus the logistic regression results will be interpretable as “probability that bird actively highlights a certain color”. (Note that this is not a problem for the highlighted-region analysis because all birds have a face, bill, underwings, breast etc.)

IV. Discussion
I will not get into much detail about the discussion because I would ideally like to see the earlier points resolved before I can turn my eyes to interpretation of the results. But I would like to highlight two points. First, I like the line of reasoning that led the authors to suggest weapons and size augmentation as universal signals. I would be curious to see the result of the new color analysis to see if it follows the same logic (is there a universal color signal?). My only suggestion here is to be more cautious about interpretation of the “closest region” results. In a neutral position, bills are always the closest point to anything birds are facing because they are the most anterior part of the avian body. So, the fact that bills were the part of the body closest to the opponent in the majority of cases can simply mean that the signaling individuals were facing the opponent, rather than actively displaying their bills. Again, I do like the line of reasoning here, but I would appreciate if the authors acknowledged this caveat. My second comment is that paragraphs #5 and 6 (L327–343) are only stating the results in different words, rather than discussing them.

V. Line-by-line comments
L99: could the authors please be more specific about when they considered that a bird was retreating from a resource and why this is relevant for data collection? Perhaps also providing an example?

L146: what was the search strategy for intraspecific signals? Did the authors direct searchers at species for which they already had interspecific data? Did they stop searching after finding a certain number of examples?

L150: could the authors please elaborate on how uniform coloration may obscure differences between intra and interspecific signals? It seems to me that, since the signaling species would be the same, coloration is automatically controlled for in pairwise comparisons between intra and interspecific signals.

L182 and 190: random effect was the same for both GLMM analyses (“Number” variable in the dataset), but described with different terms in the text. I would suggest using the same term for clarity.

L364: it would be interesting to see examples of studies that omit these regions.

L367: I would ask the authors to be more specific on how more subtle differences between intra and interspecific signals could not be picked up by their study.


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

Input:
Title: AGGRESSIVE SIGNALING AMONG COMPETING SPECIES OF BIRDS
Review round: 1
Reviewer: 2

Output:
Basic reporting: This paper and its presentation are well-structured, and contains useful figures/tables and also the raw data and code. The introduction is well-presented, but the paper could benefit from a few minor word choice changes to improve clarity, as well as a key word choice change: I encourage the authors to think more carefully about the use of the word “highlighted” and to perhaps come up with a different way to refer to the regions of the body/colors that birds displayed during aggressive interactions.

-	Minor wording suggestion: I suggest changing final line of intro (84-87) from “For groups with sufficient sample sizes, we also used this evidence to compare postures and body regions used by different individuals (3) within species, and (4) among taxonomic families of birds, looking for consistencies and differences.” to “For groups with sufficient sample sizes, we also used this evidence to assess variation in postures and body regions used by different individuals (3) within species, and (4) among taxonomic families of birds.”
-	In Line 104, the placement of the Gill et al 2020 citation struck me as a bit odd, but it looks like you included that citation to identify the taxonomy you followed for species identification. In that case, you should say that, e.g. “…and 50 different families, following the IOC World Bird List taxonomy (Gill et al 2020).”
-	Line 106-109 are redundant with the sections that follow.
-	In the “Highlighted body regions” section, I think it is important that you define what “highlighted” means early in the paragraph (e.g. before you discuss grouping of regions).
-	Line 123: what does “distinguished” mean here (and who is doing the distinguishing)? Do you mean that the authors of studies of coloration paid special attention to those regions, or that birds in those studies distinguished those regions somehow?
-	In Line 138, does “focal regions of signaling” mean the same thing as “highlighted regions”? If so, please use consistent terminology. I prefer the “focal regions of signaling” terminology, since “highlighted” is passive and makes the actor (who is doing the highlighting, the bird or the authors?) unclear.
-	Line 139, the use of the word “highlighted” is a bit unclear. I think you are saying that the displaying bird is “highlighting” regions that are particular colors, in the direction of the other individual. Please clarify: Is the protocol of your naïve scorers to examine a video/photo and identify three body parts that are prominently featured in the bird’s display and oriented toward another individual, and then to say the dominant color of the three body parts? Or do the naïve observers identify conspicuous colors/color patterns that are not necessarily on the three “highlighted” body parts?
-	The sentence starting on line 185 should be the start of a new paragraph.
-	In addition to the cases mentioned above, there are several sentences in the manuscript that use passive voice, which should be changed to active voice. For example, line 226 says “no one shoulder position was taken in the majority of aggressive signals…”
-	Line 375 do you mean auditory signals, not visual?
-	Word choice: line 379 you say your study highlights but also that there are highlighted regions. This goes back to my sense that “highlighted” regions or colors is a confusing word choice. In any case, the wording in this line should be altered. Also, I suggest using “differences” instead of “distinctions” in line 380.
-	Line 326 needs a citation for bird-mammal signaling
Experimental design: The authors have clearly contextualized the work and addressed a knowledge gap. The data they collected are relevant for answering their outlined question and the analysis is done carefully. However, the paper requires additional details in the methods section. Furthermore, I have several significant questions and suggestions about the analysis.

Methods:
-	Line 93 says that most of the videos were from WikiAves. That implies that other databases were used; please list these as well. In addition, please explain how you found videos/photos from these other databases (where did you search for them, what search terms did you use).
-	Related to that, Figure 9 is a photo from Cornell bird cams (and is a great depiction of interspecific aggressive signaling!), which suggests that you might have analyzed some of the feeder camera footage? Is that the case? If so, please say that in the figure caption.
-	Please include a short description of WikiAves for readers who are not familiar with the database (for example, the geographic range covered by this database, who contribute data to the site, what years of observations does the database cover).
-	Please share more information about how you searched for photos/videos in WikiAves. There is an Advanced Search function in the database; did you use this, and if so, which search criteria did you use? (e.g., what did you select for “Photo content”, for “Main Action”? Did you examine all photos/videos that came up from this search or only particular dates?)
-	In Line 103-104 (and later in the discussion), please also include the number of orders (not just genera and families).
-	The “Postures,” “Highlighted body regions,” and “Highlighted colors” sections all refer to only interspecific interactions, and the “Relationship to intraspecific signals” section indicates that the photos/videos you analyzed were interspecific not intraspecific interactions. Therefore you should say in the first section, “Signal data”, that the photos/videos you collected were about interspecific interactions.
-	That being said, I think the fact that some of the data on intraspecific signals were based on literature descriptions and thus collected in a different way than the interspecific signals info is a potential problem. The authors can address this by verifying that the video/photo approach yields comparable information to the descriptions in the literature. Please use the same protocol of examining/categorizing signaling in video/photos of intraspecific interactions for a subset of species and compare those findings to the intraspecific signaling descriptions from the literature.
-	Lines 152-154: please say for how many species you used literature descriptions and for how many species you used videos/photos.

Analysis:
-	It seems to me that your GLMMs should include signaling species and/or signaling family as a random effect. Also relevant to include as a random effect is whether the data come from a photo or a video (for example, it seems that an observer would be more likely to identify an open beak as important in a video where a bird is opening and closing its beak (since any opening of the beak during the clip would count as an open beak), as opposed to a photo where the image might have been taken right after the bird closed its beak).
-	While the conditional inference trees (CIT) are an interesting way to present the data, I am not convinced that they are the appropriate analysis and visualization option here and would like the authors to provide some explanation about why they chose the method. I am familiar with classification and regression trees (CART), random forests, etc., and understand the difference in splitting criterion between CIT and CART. It seems to me, however, that CIT must suffer from one of the same limitations as CART, which is that the model is built by making univariate splits after linearly ranking predictor variables, when in fact the “true” relationship between predictors and the response could be nonlinear. As such, a classification trees (and perhaps CIT) suffer from much weaker predictive ability (especially when not pruned) than random forests, in exchange for being easier to interpret. So, I think the question here is—are you building a CIT to describe variation within and between families, or to build a model that generally captures important predictors of family-specific behavior (and thus want your model to do a good job of categorizing species)? If your goal is the latter, a random forest approach (using a CIT-type algorithm) would definitely be better. However, I think your goal is the first. I think your description in the results section of the CIT models you generated are consistent with this modeling approach (e.g., the most important variable is mouth, line 270), but in the discussion it sounds more like you are commenting on variation in and among families. For example, in the paragraph starting on line 327, you comment on certain postures or body regions used by certain families, as opposed to the CIT category which combines multiple postures or body regions. For this reason, I don’t think a CIT model is best. I feel that simply providing descriptive statistics and a visualization such as a grid of family x posture (or family x body region, or family x color) would be much easier to interpret and more in line with the goal of identifying commonalities and differences among families as well as variation within families. If you disagree with my assessment of the use of CIT, please provide more justification and explanation in the methods section.
-	Line 201-202: You state that the goal of the CITs is to “examine how well taxonomic families could be categorized based on their postures,” but this wording is unclear (do you mean to say that you want to know whether different species in a family are placed into the same or different categories, or whether different taxonomic families could be grouped together?). Also, I think this stated goal follows the goal of CITs, but as described in the previous comment, I am not sure that this goal aligns with your previously stated goal of characterizing family-level variation in aggressive signaling (e.g., Line 86-87).
-	If a certain family never highlighted certain colors (for example, carotenoids), is that because they have such coloration and don’t highlight it, or might it also be because they lack such coloration?
Validity of the findings: The authors have discussed the implications clearly without overstating the work. I found the points they brought up in the discussion to be interesting and well expressed. I have a few line comments for the discussion as well as some suggestions for additional topics to elaborate on in the discussion.

-	Line 293 – I think you could cite only Fig 3. I have trouble easily gleaning this result from Fig 6 and 8.
-	Line 294-295 Isn’t the result that the bill was oriented forward (either open or closed)? Or does forward mean close to the receiver?
-	Discrepancy between sentence on line 338-339 and line 339-342, wording implies that black/white are carotenoid colors.
-	Line 364-366: do you mean something like: “Importantly, we find that certain areas that have been omitted from studies of aggressive signaling, such as bright mouth linings and underwing patterns, are in fact frequently emphasized in the aggressive displays of many bird species.”

Suggestions for additional discussion:
-	Discussion should discuss taxonomic/geographic limitations of your study. Do you expect any other kinds of signaling or any deviances from the patterns you found if you were to include other major groups of birds, or do you think these results can be broadly applied?
-	Also, please discuss how aggressive signals might vary in other contexts. You didn’t examine cases where the birds ended up fighting or where one retreated; would you expect different signals to be used prior to those fights or in those situations?
-	Any comments about WikiAves as a dataset? Or the value of citizen science databases like it?
Additional comments: Overall, I found the paper to be straightforward and a nice use of an interesting database to address a gap in our knowledge of interspecific aggressive interactions. I am confident that the authors will be able to address my comments.

I have a few remaining line comments:
-	Figs 1 and 8: beautiful drawings! Please cite the artist (even if it’s one of the authors).
-	Fig 3 and 4: can you jitter the gray points vertically or say the sample size at 1 or 0 instead? Currently it’s hard to get a sense for how many points are on the plot.
-	Fig 5,6,7 remove node numbers so we focus on sample size numbers instead
-	Small typo in the R file: you refer to the column signal.dat$signalling.families but in the dataset you provided, the column is called signaling.families.


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: AGGRESSIVE SIGNALING AMONG COMPETING SPECIES OF BIRDS
Review round: 2
Reviewer: 1

Output:
Basic reporting: No comment.
Experimental design: No comment.
Validity of the findings: Although the new analytical approach is undoubtedly more robust, there is still an issue with the PGLMMs (see Additional comments).
Additional comments: I appreciate the modifications made by the authors in response to my previous comments. They made substantial changes to address all of my major concerns. I am happy with the clarification about what I suspected could be circular reasoning in data collection and scoring. I was also glad to see that the authors rescored color data so as to make the color analysis more biologically relevant. The manuscript overall also reads more focused and streamlined in this version, in part because one of the previous analyses was appropriately removed. Finally, the new, phylogenetically controlled analytical approach provides much more robustness to the authors’ findings (but see caveat below). Overall, I find the manuscript in its new version to be a valuable contribution to the avian aggressive signaling literature.

I have a few minor comments that I will outline below.

L153: “We defined highlighted regions as those that were more obviously displayed or modified for display to the receiver” – seeing as naïve observers, not the authors, were responsible for scoring highlighted regions, it seems to me that the definition should be whatever was passed as instructions to the observers. No definition was provided to observers according to Appendix II, in which case I would suggest that this sentence (L153–155) be excluded. The next sentence (“perceived as most highlighted [by observers]”) more accurately describes the scoring procedure.

L158 and 169: reference to Appendix II would help readers.

L215: I would hate to have the authors conduct these analyses once again, but, as I had pointed out in my previous review, the best practice for PGLMMs with multiple measurements is to include species identity as a random effect, *in addition* to phylogeny. See http://www.mpcm-evolution.com/practice/online-practical-material-chapter-11/chapter-11-2-multiple-measurements-model-mcmcglmm.

L355–371: I like this paragraph relating aggressive displays to weapons. Perhaps the authors could also comment on the species of birds that possess specialized weapons (bony spurs) on the wings or leg, two of which are in the dataset (Vanellus and Jacana; also Gallus but not hens)? Spurs appear to be highlighted by both species in the assessed pictures.

By the way, wing pounding is not unique to pigeons and doves (L370). Besides Columbiformes (e.g., account by Leguat 1708 in Hume & Steel 2013, https://doi.org/10.1111/bij.12087), species in Anseriformes (e.g., Nuechterlein & Storer paper cited in the manuscript) and Charadriiformes (e.g., Walters 1979, https://doi.org/10.1016/0003-3472(79)90045-9) that possess wing spurs use them for wing pounding.

L374: Body size is also a good predictor of winner of fights across animals (Palaoro & Peixoto 2021, https://doi.org/10.1101/2020.08.26.268185).

L452–459: I really liked the authors’ decision to include this paragraph in the main text rather than in the acknowledgments as would be expected. I was glad to see this invaluable resource from my home country duly emphasized.

Figure 2 looks so much nicer now, with the estimates and 95% CI instead of the bar plots. Two minor suggestions: sort panels (c) and (h) like the other panels; indicate in panel (c) what positions were grouped as “open wings”.

Figures 4 and 5: I suggest reversing the time axis (i.e., starting at ~120). The axis would then be interpretable as million years before present (Mybp) or million years ago (Mya) — more intuitive to readers than branch length.

Love new Figure 5 — very informative! I would consider using black-and-white striped pattern for dark/white contrast and white tile with black borders for white (not sure if the latter will work well, but I think it is worth trying). Also, I think the authors meant “brown for brown.beige” in the legend.


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

Input:
Title: AGGRESSIVE SIGNALING AMONG COMPETING SPECIES OF BIRDS
Review round: 2
Reviewer: 2

Output:
Basic reporting: The authors have addressed many of my concerns and have greatly improved the manuscript. However, I still found parts of the manuscript difficult to understand and think that the authors should revise to improve clarity, especially in the methods. I also have major concerns about the choice of statistical analyses.

-	Lines 104-109: Please clarify this wording. From these two sentences, it’s unclear whether the data from the Internet Bird Collection ended up being used or not, given that you did not include data from the Macaulay Library. Do you mean that OTHER data from the Macaulay Library (not the IBC data that you had searched before it was moved to the Macaulay Library) you didn’t use, but you did use IBC data? It’s unclear when you say “we omitted this data source from our study” which data source you are referring to.
-	On the topic of the choice of the word “highlighted”, I agree with the authors that it is important to use that terminology given that that was the term they used in instructions to data collectors. However, please avoid using the passive voice (e.g., “were highlighted” in Line 136) when you do use this word, to improve clarity.
-	Lines 210-212 make it sound like the postures/colors analyses were done for intra vs interspecific comparisons. Consider rephrasing to something like: “We used phylogenetic mixed-effects models using the MCMCglmm function in the MCMCglmm R package (version 2.32; Hadfield 2010) to test whether birds were more likely to use specific postures or highlight specific body regions or colors in aggressive encounters with heterospecifics, as well as to compare whether birds use the same signals in intraspecific aggressive interactions as in interspecific aggressive encounters.”
-	I’m very confused by 244-249 wording. What does it mean for a bird to assume a degree of similarity?
-	Line 259 suggested wording change: “we calculated the proportions”
-	Figure 2 and Figure 3b, 3c are low quality/blurry and difficult to read. Please improve the quality and make the text larger/points darker to improve legibility.
Experimental design: -	The wording of line 189-191 is confusing. First, I don’t think you mean that species are uninformative, so you should write something like: “We did not incorporate color into these comparisons because some species have uniform coloration and thus color is uninformative with respect to patterns of within- versus among-species signaling”. Second, it doesn’t make sense to ask if a bird “highlights” a color when it has a uniform color/is only that color, regardless of whether you’re doing within- or between-species comparisons. Please clarify if you are including birds that are only one color in your color analyses. Do you instead here mean “…some species have no/low within-species color variation…?”
Validity of the findings: -	The interpretation of the glms starting on Line 270 is very confusing. I had trouble matching the p-values reported in the text with the values reported in the supplemental tables. I also don’t agree that it is okay to interpret an intercept with a p-value > 0.5 as being a significant driver of the outcome just because the other predictors in the model are significantly lower than the intercept. Most importantly, however, I do not think that models in the GLMM family are appropriate for this analysis (and for some of your other analyses). If your question is whether or not the different options for a given posture, e.g. body position, are equally likely, I think you want a model in the chi-square family. This same critique can be applied to many of your analyses. I believe that the authors need to re-examine and simplify many of their analyses and work on the description of their analysis to make what they did very clear to the reader. If I have misunderstood, please explain and justify your methods better.
-	In your supplemental tables, please label the p-value as pMCMC, and in the table caption clarify that the 95% CI are credible intervals.
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: AGGRESSIVE SIGNALING AMONG COMPETING SPECIES OF BIRDS
Review round: 3
Reviewer: 1

Output:
Basic reporting: See below.
Experimental design: See below.
Validity of the findings: See below.
Additional comments: I appreciate the authors addressing the few comments that I had made on my previous report and maintain that this manuscript would be a valuable contribution to the aggressive signaling literature.

I do have to say that, upon inspecting the codes, I came across issues that I had failed to notice in my previous revision. I apologize for bringing up new topics in the third round of revision but hope that the authors understand that refraining from doing so would mean not serving appropriately in my role as a reviewer. Most importantly, however, I trust that these issues (a) will not qualitatively (and barely quantitatively) affect the results and (b) will not be hard to deal with.

The first is what is likely a typo in the codes. All priors were defined as lists containing the element “n”, when MCMCglmm needs the element “nu”. Simply replacing all instances where “n=k” occur with “nu=k” solves this issue. I am not completely sure that this is indeed a typo — perhaps it is an issue that arose in the new version of MCMCglmm (the authors used v2.32 for analyses, while I am testing their codes with v2.33).

The second is that the species Torgos tracheliotos appears with two different IOC codes (signaling.IOC) in the data set, causing this species to behave differently in relation to the random factors “animal” (such that they are considered to be on the same tip of the phylogeny) and “signaling.IOC” (such that they are treated as distinct species). I corrected this issue (i.e., made it so that all Torgos tracheliotos have the same signaling.IOC) and reran all models with much fewer iterations as a test. Results changed only very slightly (if at all). I am pretty sure that correcting this issue will barely change the results quantitatively (certainly not change them qualitatively) and so will leave it to the editor’s and authors’ discretion to decide on whether it is worth the trouble of rerunning analyses.

The third is related to the authors’ choice of using no thinning interval for the MCMC chains. Using no thinning with traditional MCMC samplers like the ones MCMCglmm uses (as opposed to more efficient samplers like HMC or NUTS) causes a very high autocorrelation between successive samples — this can be verified with the function coda::autocorr. Excessive autocorrelation is not a problem in itself but it invariably leads to very reduced effective sample sizes (ESS). That is why ESS rarely exceeds 30,000 in Table S12, even though each chain sampled 990,000 estimates per parameter. The authors mention in the Supporting Information that an ESS of 200 is satisfactory*. I could not find this information in the references provided and would appreciate if the authors could point me to it, because I seem to remember seeing higher cutoffs generally being used as a rule of thumb (at least 1000, if I am not mistaken). Not all parameters would pass a 1000 cutoff, especially for the model in Table S12i. For future reference, I would recommend that the authors use a thinning interval of at least 100 — this will improve the ESS-to-total sample ratio and spend less memory on redundant samples. But for the purpose of this study, there is an easier way around that does not require rerunning models.

[*By the way, the authors mention that an ESS > 200 denotes chain convergence, but I find this statement to be strange because, as far as I know, ESS provides no information on chain convergence. ESS is simply an estimate of how large the sample would be if estimates were independent from each other. This information can indicate how efficient the sampler is, not whether chains converged well. Convergence, again as far as I am aware, is instead tested by comparing the behavior of multiple chains, like Gelman–Rubin diagnostics do.]

Since the authors ran three chains per model to test convergence, and they found that the chains did converge, the solution is simple: they can safely combine the estimates from all three chains (which they already have in hand — no need to rerun) and thereby increase ESS by a factor of approximately three. Regardless of cutoffs, a higher ESS will always provide more robust estimates, and that is especially true for Table S12i. To implement what I am suggesting, all the authors have to do is combine the chains before computing ESS, mean, and 95% CI. I will provide an example for the body orientation model below — hopefully it will be of help for the other models as well.

# combining fixed effect estimates as a list of matrices
Sol <- mcmc.list(body.orientation.model.MCMCglmm$Sol, body.orientation.model.MCMCglmm.2$Sol, body.orientation.model.MCMCglmm.3$Sol)
# extracting ESS
round(sort(effectiveSize(Sol)))

# combining random effect estimates
vv <- mcmc.list(body.orientation.model.MCMCglmm$VCV, body.orientation.model.MCMCglmm.2$VCV, body.orientation.model.MCMCglmm.3$VCV)
# extracting ESS
effectiveSize(vv)

# combining fixed effect estimates as a single matrix
est <- rbind(body.orientation.model.MCMCglmm$Sol, body.orientation.model.MCMCglmm.2$Sol, body.orientation.model.MCMCglmm.3$Sol)
# extracting mean for forward.lowered
forward.lowered.est<-inv.logit(mean(est[,"(Intercept)"]))
# extracting 95% CI for forward.lowered
forward.lowered.ci<-inv.logit(HPDinterval(mcmc(est[,"(Intercept)"])))

Finally, a small correction regarding L140: spurs in the mentioned species are located in the carpal joint (i.e., the wrist, not the elbow).


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: EFFECTS OF TURNING OVER INTENSITY ON FATTY ACID METABOLITES IN POSTHARVEST LEAVES OF TIEGUANYIN OOLONG TEA (CAMELLIA SINENSIS)
Review round: 1
Reviewer: 1

Output:
Basic reporting: 1. There are some grammatical mistakes. It is suggested that we should standardize our own words, I strongly recommend that the authors should check the manuscript more thoroughly and ask fluent English speaker to edit their manuscript.

2. The format is chaotic, either texts or reference. For instance, non-uniform units, non-standard Latin names, casual typeset, and un-regulated superscript and subscript. Please check them through the manuscript.

3. Introduction section should give more details for readers. Especially, give more details of new advances in research on fatty acids in Camellia sinensis. The literature regarding LOX genes cited in the introduction is older, please add more relevant literatures support for the past three years.
Experimental design: 1. Materials & Methods section Is there any reference or pre-test for this treatment method, that is, whether this method is reasonable or not, please add reference or pre-test data.

2. Statistical analysis is missing. It needs to be supplemented in Figure 3A.

3. The image of Figure 3C is blurred and can't be seen clearly. It is suggested to adjust and modify Figure 3C and other unclear pictures.
Validity of the findings: 1. The results of LOX genes in Camellia sinensis are inconsistent. According to the table 1, 13 LOX genes were mined, however, in Figure 4 and 5, only 12 LOX genes were investigated. where is CsLOX13? These discrepant results should be better illustrated.

2. The authors declared that “CsLOX1 might be the main gene to play an important role in the degradation of fatty acid substrates and the performance of key fatty acid derived volatiles”. It is an interesting speculation. More detailed evidences are required.

3. Discussion section “Techniques in investigating volatiles” is incomplete and deficient. More information is required. Furthermore, what is the link between “Techniques in investigating volatiles” and other topics?
Additional comments: Manuscript ID: 68064
Manuscript title: Effects of turning over intensity on fatty acid metabolites in postharvest leaves of ‘Tieguanyin’ oolong tea (Camellia sinensis)

In present manuscript, the authors investigated the effects of turning over intensity on fatty acid metabolites, including total fatty acids, saturated fatty acids, unsaturated fatty acids, and characteristic fatty acid derived volatiles, in postharvest leaves of ‘Tieguanyin’ oolong tea (Camellia sinensis). Overall, it has brought some interesting results. However, the following concerning should be also addressed.

The results of LOX genes in Camellia sinensis are inconsistent. According to the table 1, 13 LOX genes were mined, however, in Figure 4 and 5, only 12 LOX genes were investigated. where is CsLOX13? These discrepant results should be better illustrated.

The format is chaotic, either texts or reference. For instance, non-uniform units, non-standard Latin names, casual typeset, and un-regulated superscript and subscript. Please check them through the manuscript.

Statistical analysis is missing. It needs to be supplemented in Figure 3A.

Introduction section should give more details for readers. Especially, give more details of new advances in research on fatty acids in Camellia sinensis. The literature regarding LOX genes cited in the introduction is older, please add more relevant literatures support for the past three years.

Materials & Methods section Is there any reference or pre-test for this treatment method, that is, whether this method is reasonable or not, please add reference or pre-test data.

The image of Figure 3C is blurred and can't be seen clearly. It is suggested to adjust and modify Figure 3C and other unclear pictures.

Discussion section “Techniques in investigating volatiles” is incomplete and deficient. More information is required. Furthermore, what is the link between “Techniques in investigating volatiles” and other topics?

The authors declared that “CsLOX1 might be the main gene to play an important role in the degradation of fatty acid substrates and the performance of key fatty acid derived volatiles”. It is an interesting speculation. More detailed evidences are required.


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: EFFECTS OF TURNING OVER INTENSITY ON FATTY ACID METABOLITES IN POSTHARVEST LEAVES OF TIEGUANYIN OOLONG TEA (CAMELLIA SINENSIS)
Review round: 1
Reviewer: 2

Output:
Basic reporting: Although the fatty acid derived volatiles are one types of key aroma compounds in oolong tea and had been well studied, in this manuscript, Ni and co-authors detected the changes of fatty acids, their derived volatiles and LOX genes’ expression patterns during oolong tea’s turn over processing. The experiments are well conducted and interpreted. The results of this study will provide a valuable information to understand the aroma formation during oolong tea processing.
Experimental design: 1.	How to perform the turn over processing?
2.	How to define the turn over intensity? From the methods, T1, T5 and T9 is the different time points during oolong tea, they have no relationship with intensity?   So I suggest the authors to change the title and “intensity” using “degree/stage of turn over processing”.
3.	How to give the names of CsLOX genes, the reference should be listed.
Validity of the findings: 4.	Line 302-310, delete.
5.	The name of genes should be expressed as italic, such as in the title of Fig. 4.
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: EFFECTS OF TURNING OVER INTENSITY ON FATTY ACID METABOLITES IN POSTHARVEST LEAVES OF TIEGUANYIN OOLONG TEA (CAMELLIA SINENSIS)
Review round: 2
Reviewer: 1

Output:
Basic reporting: The study provides an overall view of how fatty acid metabolites change and affect the quality of oolong tea with different turning over intensities during processing. The manuscript is well-written after the revision.
Experimental design: The Experimental Design of the manuscript is reasonable and the method is described in detail.
Validity of the findings: The fingdings are of great significance in guiding the control of tea aroma quality.
Additional comments: The manuscript is suggested to be accepted for publication in the 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: IDENTIFICATION AND EXPRESSION ANALYSIS OF XYLOGLUCAN ENDOTRANSGLUCOSYLASE/HYDROLASE (XTH) FAMILY IN GRAPEVINE (VITIS VINIFERA L.)
Review round: 1
Reviewer: 1

Output:
Basic reporting: The manuscript "Identification and expression analysis of xyloglucan endotransglucosylase/hydrolase (XTH) family genes in grapevine (Vitis vinifera L.)" reports the genome-wide identification of XTH genes and expression profiling in grapes.
XTH genes are known to play important role in plant development as well as response to abiotic and biotic stresses. Therefore, this MS will be of interest to grape researchers.
The manuscript is well written. Sufficient background information is given.
Experimental design: The experimental design is appropriate. The methodology is given in sufficient detail.
The results have been given in detail covering all the aspects. All the relevant figures and tables have been included, the figure quality is good. The relevant raw data has been shared.
Validity of the findings: The conclusions are well stated and linked to the original research question.
Additional comments: No additional comments