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: EMBRYO AND LARVAL BIOLOGY OF THE DEEP-SEA OCTOCORAL DENTOMURICEA AFF. METEOR UNDER DIFFERENT TEMPERATURE REGIMES
Review round: 1
Reviewer: 3

Output:
Basic reporting: See comments in General comments for the authors
Experimental design: See comments in General comments for the authors
Validity of the findings: See comments in General comments for the authors
Additional comments: The manuscript of Dr. Maria Rakka et al. deals with a deep-sea octocoral and emphasizes the little knowledge existing on their early life history stages. The study describes the embryo and larval biology of Dentomuricea aff. meteor which is common species in the Azores. The experimental work was conducted under two temperature regimes, corresponding to the minima and maxima existing in their natural habitat during the reproductive season.
See comments below:
1.	Indeed, the life history and reproductive biology of deep sea octocorals is less known compare to the shallow-water species. In this respect the body of information presented in the manuscript is novel and interesting.  However, there are some flaws in the narrative of the manuscript which are addressed below.
2.	The authors present the goal of the text (lines 83-88), but I would rather expect reading a clear working hypothesis with some specific research questions. Otherwise, the current narrative weakens the text and mainly the interpretation of the results. In its current format the manuscript is a kind of a descriptive report which can be acceptable, but I think it does not fit Peer J. As correctly stated by the authors, temperature is a critical environmental factor in the marine environment which greatly affects coral reproduction.  The most intriguing question that comes up while reading the ms is what are the effects of ambient seasonal temperature changes on the early developmental stages and life history of D. aff. meteor. However, it is not clear what is the relevance of global change, in particular global warming to the present study, since it deals with natural and normal temp. fluctuations in the deep sea environment and do not present evidence for temp. change there.
3.	A general comment: throughout the text where average± SD values are presented, sample size (n value) should be given (e.g. line 27, 29, 264, 270, etc.)
4.	Lines 102-107 describe the conditions in which the octocoral fragments were maintained and indicate that the seawater was pumped from 5 m. Has the water quality in terms of pH, nutrients, salinity, etc. been compared to the seamount ones? If I read correctly, only the temperature was well controlled. Similarly, it sees that the filtration and UV treatments were applied in order to remove any microorganisms from the water and making them sterile. Did the author use any control to test possible effect of these procedures on the octocoral fragments?
5.	The materials and methods section dealing with the embryonic development, larval studies, and their settlement and metamorphosis. I couldn’t find an indication which type of water was used for those assays. Did the authors use the same filtered/UV treated seawater as for rearing the fragments?
6.	The Results section that deals with the spawning activity (line 229 onward) mentions that it has not been possible to directly observe fertilization and to differentiate between the possibility of internal vs. external fertilization. It should be noted that SEM examination, could have answer this question by looking for sperm in the polyp cavities or the oocyte surface,
7.	I have some doubts concerning the results presented in the settlement and metamorphosis section (line 303 onward). The authors did not indicate if the basalt substrate was sterilized or had some organic film on it. Since it is well established in the literature that organic film may induce settlement and metamorphosis, this parameter should be considered while running those experiments. Similarly, there is no indication for the substrate texture and whether it played any role in enhancing settlement. Indeed, settlement took place also on the glass but the settlement rate must be normalized per substrate-type before drawing conclusions.
8.	Figs. 2 & 4 present percentages and Fig. 3 presents probabilities, but sample size (n values) are missing which makes it hard to consider the strength of the results.
9.	The discussion seems to me too long and some paragraphs are wordy (e.g. line 433 onward). For the sake of clarity, I recommend to present a comparative table with the different reproductive/life history traits of the studied species in comparison to previously studied deep sea octocorals, or even some shallow water ones. For the latter, I wonder why the authors did not cite the only existing review paper on octocoral reproduction by Kahng et al. 2011.
10.	The Conclusion section (line 480 onward) should be rewritten to reflect both the novelty of the results and their general significance. For example, the sentence (line 489): “However, species distribution is a result of complex interactions between factors in various ecological levels” includes two vague notions (in bold). Those two should be clearly explained.

.


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: EMBRYO AND LARVAL BIOLOGY OF THE DEEP-SEA OCTOCORAL DENTOMURICEA AFF. METEOR UNDER DIFFERENT TEMPERATURE REGIMES
Review round: 2
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The manuscript has been appreciably improved. I am satisfied with authors' corrections and answers. However, before publication, the authors would need to still address some little corrections. I also include some suggestions that I hope can be useful. Finally, I would like to congratulate the authors for the magnific work done. Working with coral larvae is not easy and, sometimes, it is very stressful. Good job!


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

Input:
Title: HIGH GENETIC DIVERSITY AND MIXING OF COASTAL HORSESHOE CRABS (TACHYPLEUS GIGAS) ACROSS MAJOR HABITATS IN SUNDALAND, INDONESIA
Review round: 1
Reviewer: 1

Output:
Basic reporting: The English prose needs revision - I have marked the manuscript to aid in that task.
The paper lacks citations to relevant studies, which I provide in my review and in a manuscript that I will share.
Some of the inferences must be reconsidered along the lines of my comments below.
Experimental design: The study involves a limited sample analyzed at one mitochondrial locus. There may be a possibility to expand the geographic scale of analysis by inclusion of results of an earlier study as described below. The methods are generally laid out in sufficient detail that subsequent workers can build upon this study.
Validity of the findings: While generally sound, some of the inferences must be reconsidered as described below.
Additional comments: Tachylpelus gigas is one of four horseshoe crab species worldwide. Knowledge of its population genetic structure would inform its management and conservation. Aini et al. collected 91 crabs from six sites in Indonesia, sequenced the AT-rich region of its mitochondrial DNA molecule, and conducted population genetic analyses. The results contribute to our knowledge of population genetic and stock structure of the species. Yet, this study is limited in some critical ways. Sample sizes are small. Just one mitochondrial locus is considered, but no nuclear loci, limiting the scope of inference. The geographic scope of coverage is limited, though it many be possible to combine these results with existing information from another study, as I discuss below. In context below, I will mention several issues with interpretation of biology and analytic results, as well as consideration of all relevant literature. The English prose can be strengthened – I have marked the manuscript to guide revision.
     Abstract. – The authors write that the genetic structure of horseshoe crabs is unchanged over millions of years. We don’t know that. I think the authors intend to write that its overall morphology is fundamentally unchanged over hundreds of millions of years. In any case, the sentence is not really needed and can be deleted.
     The last sentence of the abstract suggests that adult and juvenile horseshoe crabs exhibit little movement among locations. In the paper, they cite a few studies of another horseshoe crab species, but not this species. To really defend the assertion for T. gigas, they’d have to cite results of long-term mark-recapture studies. Populations genetics indexes movement over recent generations, and mitochondrial studies index only female movement. Males being smaller, they tend to move greater distances. My comments here indicate that this sentence must be carefully revised to prove defensible.
     Introduction. – At line 55, I think that the word “movement” may be missing.
     At line 66, the issue of genetic structure being unchanged for millions of years arises again.
     At line 61, Limulus polyphemus is more frequently referred to as the American horseshoe crab.
     At line 69 is a sentence on genetics that seems out of place. I suggest it be moved to line 77. The passage fails to mention two more recent studies of relevance, King et al. (2005) and (2015). Further, an existing study of population genetics of T. gigas by Roihan and Ismail (2011) is relevant in this context, but is not mentioned.
     Methods. – It seems that different methods of calculating Fst were employed. Using DNA sequences, Fst as calculated by Rozas et al. (2017) would be preferred. Fst from AMOVA analyses is less precise.
     Results. – At lines 166-169, negative values of Fst are not biologically meaningful, they are to be interpreted as essentially zero.
     At line 169 and other contexts, geneflow Nm should be reported with the unit of individuals per generation.
     At line 176, the authors should not write of “dominant”, but rather “common” haplotypes. Dominance is a descriptor relevant to Mendelian traits.
     At line 177, the authors should write that shared haplotypes are an indication of gene flow among geographic sites. Writing of evolutionary links suggests an indication of phylogenetic relationships, which were not considered in this study.
     In the last paragraph of Results, the authors write of historical demography and seem to regard their results from the DNA sequence mismatch analysis and from the structure of the haplotype network as conflicting. This is not necessarily so; the DNA sequence mismatch speaks to deeper time, to a demographic expansion perhaps coincident with the post-Pleistocene rise of sea level. The haplotype network structure speaks to more recent demographic processes. I must add though, that small sample size limits the possibility of observing the intermediate haplotypes that are inferred to exist on the network.
     Both this study and Roihan and Ismail looked at variation at the same mitochondrial region. If the earlier data are archived, then the authors can combine the existing and new data, trim the sequences to the same length, and repeat the analyses to thereby gain a view of horseshoe crab population genetic structure over a much larger area. This would contribute much to the study.
     Discussion. – At line 205, the authors compare the population genetic variation in their study with that observed by Roihan and Ismail – are the respective studies of similar scale? Is this a viable comparison?
     At line 212, the authors write that observation of a large number of haplotypes in their data indicates a high mutation rate for mitochondrial genes. This is not a suitable explanation. It is rather that the populations studied are sufficiently large to maintain a high level of genetic diversity, i.e., haplotypes are not being lost to random genetic drift.
     Throughout the discussion, the authors do not adequately consider the importance of adult dispersal. There are long-term tagging studies that show that the males in particular can move hundreds of kilometers over their long lifetimes. I will include a manuscript in review – which can be cited as a personal communication - to indicate that. This might be cited at lines 233 and 257.
     At line 139, the authors argue that inbreeding may be occurring within horseshoe crab populations. With large populations, considerable dispersal, and late maturity, this does not seem likely. High Fis metrics are more likely the result of Wahlund effect.
     At line 240, again negative Fst should be taken as Fst being effectively zero.
     At lines 250 and 267, small sample size also explains the results of this study.
     At line 253, I do not consider the haplotype network as having the classical start profile cited. It is more complex than that, so explanations would have to be more complex and nuanced.
     At line 263, Sundaland was not glaciated and deglaciated, rather it was dewatered and then inundated.
     The sentence at line 275 suggests that high mitochondrial sequence variation is adaptive.  This is oversold. It may be associated with high nuclear diversity, some of which may be adaptive, but the sentence as written is indefensible.
     At line 280, the Walls et al. reference concerns L. polyphemus, another horseshoe crab, so the extension of that assertion to T. gigas seems indefensible as written.
     The section should end with a call for a study across the range of the species and including nuclear DNA markers such as microsatellites or SNPs. I found such a call at the end of the Conclusions section. That passage should be moved up. Undiscussed material does not belong in the Conclusion.
    Conclusions. – At line 288, the two parts of the sentence do not belong together.
     References. – I marked some minor departures from journal citation stylistics.
     Tables and Figures. – The footnote to Table 1 does not seem to be cited in the label or body of the table.
Literature cited in this review:
King, TL, Eackles MS, Aunins AW, Brockmann HJ, Hallerman E, Brown BB (2015) Conservation genetics of the horseshoe crab (Limulus polyphemus): allelic diversity, zones of genetic discontinuity, and regional differentiation. Pages 65-96 in   Carmichael R, Botton ML, Shin P, Cheung SG, eds. Changing Global Perspectives on Biology, Conservation and Management of Horseshoe Crabs. Springer, Berlin
King TL, Eackles MS, Spidle AP, Brockmann HJ (2005) Regional differentiation and sex-biased dispersal among populations of the horseshoe crab, Limulus polyphemus. Trans Amer Fish Soc 134:441-465


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

Input:
Title: HIGH GENETIC DIVERSITY AND MIXING OF COASTAL HORSESHOE CRABS (TACHYPLEUS GIGAS) ACROSS MAJOR HABITATS IN SUNDALAND, INDONESIA
Review round: 1
Reviewer: 2

Output:
Basic reporting: The manuscript  requires substantial improvement and language editing
Literatures cited can be improved
Experimental design: Following the aims and scope of the journal
Further improvement in the transparency of the method adopted is to be addressed in the revised version
Validity of the findings: Authors need to argue more on the conclusion made in this study. Please refer the attached file
Additional comments: Author could refer the PDF file attached here for comments and suggestions.
Please do refer 'Mitochondrial DNA Signature for Range-Wide Populations of Bicyclus anynana Suggests a Rapid Expansion from Recent Refugia'


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

Input:
Title: HIGH GENETIC DIVERSITY AND MIXING OF COASTAL HORSESHOE CRABS (TACHYPLEUS GIGAS) ACROSS MAJOR HABITATS IN SUNDALAND, INDONESIA
Review round: 1
Reviewer: 3

Output:
Basic reporting: There is room for the improvement of English language use. Many sentences, especially ones in the introduction and discussion, are unclear. Background and literature references are provided sufficiently. The manuscript's structure is generally good, but it is not necessary to have a single paragraph section of "The Implication for Conservation" outside of "Discussion". The manuscript is mainly a report of observation; therefore, I don't think it is necessary to have a hypothesis. It seems the authors emphasizes the genetic connectivity a lot as the major contributor to the low genetic differentiation. However, population genetic analyses with a single monoploid sequence is insufficient to infer gene flow.
Experimental design: no comment
Validity of the findings: The research is original even though the methodology used is common. The research question is not well defined in the introduction, but it doesn't matter as the results are mainly descriptive. The manuscript certainly addresses some questions and discovered that the horseshoe crabs across multiple Indonesia islands are of low genetic variance, which has not been reported before. The investigation is sound enough and the statistics are robust enough. Methods described provide enough information for replication.
Additional comments: The manuscript is important as the first report to show the shallow genetic differentiation of a marine flagship species across Indonesia. The major limitation is that the use of single genetic marker of monoploidy provide limited resolution to explore further into the causes of such shallow genetic differentiation, such as gene flow, genetic drift and other dramatic demographic processes. I would recommend the publication of the manuscript with substantial revision. Unless authors decide to use additional markers and analyses to support that the shallow genetic divergence is mainly contributed by genetic connectivity, I'd suggest to only preserve "gene flow" as a possible scenario in the discussion section.


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

Input:
Title: HIGH GENETIC DIVERSITY AND MIXING OF COASTAL HORSESHOE CRABS (TACHYPLEUS GIGAS) ACROSS MAJOR HABITATS IN SUNDALAND, INDONESIA
Review round: 2
Reviewer: 1

Output:
Basic reporting: Please see the general report below
Experimental design: Please see the general report below
Validity of the findings: Please see the general report below
Additional comments: Tachylpelus gigas is one of four horseshoe crab species worldwide. Knowledge of its population genetic structure would inform its management and conservation. Aini et al. collected 91 crabs from six sites in Indonesia, sequenced the AT-rich region of its mitochondrial DNA molecule, and conducted population genetic analyses. The results contribute to our knowledge of population genetics and stock structure of the species. This is revised manuscript, and while improved, still needs clean-up of scientific presentation and prose. I have marked the manuscript to guide revision.
     Title page. – Shouldn’t the name of IPB University be spelled out?
     Abstract. – At line 29, the sequences were amplified using PRIMERS FOR mitochondrial ….
     At line 33, the AMOVA SHOWED MOST VARIATION within (95.23%) rather than among…
     At line 37, the lack of genetic structure argues for regional, as opposed to local-based conservation. In that same line, this is a study of modest scope and so its conclusions must not be oversold – the authors should delete “conclusively”.
     Introduction. – Throughout the review of this paper, there has been a contention between myself and the authors about how much the crabs move. There have been but a few marking studies, with recaptures likely limited over space and time, that the authors repeatedly cite to argue that movement is little. Yet, longer-term studies of other horseshoe crabs and their own genetics results suggest considerable movement. Part of the issue is that genetics marks movement over a multigenerational period. The authors must distinguish between individual movement as measured by marking and inter-generational migration measured with genetic structure. This issue first comes up at line 54 where the authors write that life-history and habitat use “indicate” restricted dispersal capacity. The Sekiguchi paper cited is a marking study, hence, the sentence should be modified to write of individual movement or simply toned down to say that these factors “suggest” limited dispersal capacity.
     The Sekiguchi (1988) citation should be moved to line 60 and a repeated sentence at line 64 can be deleted.
     Methods. – At line 92, the manufacturer of the GeneAiD extraction kit should be given.
     At line 102, UV light WAS USED TO OBSERVE the band which indicates…
     At line 103, where is 1st Base located?
     The sentence at lines 117-118 explaining Fst is not needed – the reader is expected to know that. Similarly, the passage from lines 126-131 explaining the D and Fs metrics is not needed. A few words of explanation in Results tells any reader unfamiliar with the metrics what positive or negative estimates mean.
     The supporting (Excoffier and Lischer 2010) citation should appear at line 128.
     Results. – The sample sizes in this study are small, yet some genetic diversity metric are presented to four decimal places. While the software does report it, the sample sizes do not justify an implicit claim of four-place precision. The authors should report their metrics to three places (three significant places if there are leading zeros). I’ve marked some, but not all such instances on the manuscript.

     At the end of line 147, the authors should report h and pi for the BP population – those values are missing.
     At line 164, the sentence should start as: RELATIONSHIPS AMONG 34 haplotypes were ILLUSTRATED… This analysis did not identify haplotypes as written.
     At line 175, the authors should tell the reader the number of haplotypes in the UK population.
     At line 177, the historical demography was assess based on the FREQUENCIES OF mtDNA AT-rich region haplotypeS.
     Discussion. – At line 208, there is LIITLE, not no subdivision among populations. The sentence as written is not correct.
     The sentence at lines 221-223 is not relevant to the passage in which it appears and should be moved or removed.
     The sentence at lines 223-226 should end the passage on movement and connectivity. The authors might add that Fst reflects inter-population movement over mutigenerational intervals that short-term tagging studies cannot.
     The sentence at line 232 regard AMOVA is not relevant to the passage and can be removed.
     The sentence at lines 240-242 on movement of horseshoe crabs is not relevant to its passage.
      The sentence at line 249 should read: The occurrence of the many unique haplotypes is explained by the SMALL sample size AND POSSIBLY ALSO the ISOLATION during the last glacial maximum.
     At lines 261-262, the authors write that all individuals do not move far along the coastline. They have no data to support this interpretation and their genetic results suggest considerable movement among sites. Best to simply delete this sentence.
     At line 269, best to start the paragraph with High genetic diversity. The authors should not oversell their results by claiming that their findings are “conclusive”.
     References. – I marked some minor departures from journal citation stylistics. The King et al. citations are not in the right place.
     Tables and Figures. – Entries in Table 1 should not claim so much precision.
     “ns” does not appear in Table 3, so the footnote defining that can be removed.


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

Input:
Title: HIGH GENETIC DIVERSITY AND MIXING OF COASTAL HORSESHOE CRABS (TACHYPLEUS GIGAS) ACROSS MAJOR HABITATS IN SUNDALAND, INDONESIA
Review round: 2
Reviewer: 2

Output:
Basic reporting: Many comments raised on the manuscript content  is not well addressed in the revised article. Authors need to focus each points and improve the manuscript substantially.
Experimental design: Additional information needed  (Refer the review file)
Validity of the findings: Sound, Novel and timely.
Additional comments: I advice authors to send the manuscript for language editing.
Further comments can be seen in review file.


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

Input:
Title: HIGH GENETIC DIVERSITY AND MIXING OF COASTAL HORSESHOE CRABS (TACHYPLEUS GIGAS) ACROSS MAJOR HABITATS IN SUNDALAND, INDONESIA
Review round: 2
Reviewer: 3

Output:
Basic reporting: There is still room for improvement in English. I have made some suggestions directly on the manuscript.
Experimental design: no comment
Validity of the findings: Still, the authors' efforts is insufficient to suggest gene flow to be the only contributor to the low divergence of the coastal horseshoe crabs across Indonesia. I'd suggest to tone down the contribution of gene flow in both abstract and introduction. At the same time, provide alternative scenario such as lack of time for divergence from stock population during the last glacial maximum.
Additional comments: In general, the manuscript as a report for observation of low genetic diversity among Indonesia horseshoe crab populations is satisfying. However, the implication of gene flow to such population genetic structure needs further investigations. Moreover, the language should be further improved.


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

Input:
Title: HIGH GENETIC DIVERSITY AND MIXING OF COASTAL HORSESHOE CRABS (TACHYPLEUS GIGAS) ACROSS MAJOR HABITATS IN SUNDALAND, INDONESIA
Review round: 3
Reviewer: 1

Output:
Basic reporting: Needs some minor corrections as noted below, and would benefit from polishing of the technical prose as marked on the scanned, marked manuscript.
Experimental design: It's a study of limited scope, but it does open the field.
Validity of the findings: With some minor prose revisions, findings will be on the mark.
Additional comments: Knowledge of the population genetic structure of Tachylpelus gigas would inform its management and conservation. Aini et al. collected 91 crabs from six sites in Indonesia, sequenced the mitochondrial AT-rich region, and conducted population genetic analyses. The results contribute to our knowledge of population genetic and stock structure of the species. Yet, this study is limited because sample sizes are small, one mitochondrial locus but no nuclear loci are screened, and the geographic scope of coverage is limited. This is a revised manuscript and is notably improved. The English prose can be further polished by attention to the marks on the scanned manuscript. A few context-dependent comments appear below.
     Methods. – So that others can build on the methods successfully employed, manufacturers of key products should be mentioned at lines 88, 93, and 94.
     Results. – At line 148 is an apparent error – it seems to me that haplotype H3 was observed in 15 individuals, not what is written that it was observed in all but 15 individuals.
     At line 204, another apparent error – seven, not six haplotypes were shared among locations.
     The sentence at line 206 should read that relationship patterns at the mtDNA level showed little geographical structure.
     Discussion. – In the Conclusion, the authors should write that locally AND REGIONALLY based conservation management is recommended. They did make this change in the abstract, but apparently omitted it here.
     References. – I marked some minor departures from journal citation stylistics.
     Hasn’t IUCN revised its Red List assessment of the species since 1996?
     Tables and Figures. – Table 1 should include a column noting the number of observations of each haplotype.


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

Input:
Title: HIGH GENETIC DIVERSITY AND MIXING OF COASTAL HORSESHOE CRABS (TACHYPLEUS GIGAS) ACROSS MAJOR HABITATS IN SUNDALAND, INDONESIA
Review round: 3
Reviewer: 2

Output:
Basic reporting: I think the manuscript is much clearer then the previous version. I still have a few minor suggestions, especially on the discussion section.

1. The language has great improvement from the last version. However, I noticed that the authors using past tense throughout when reporting methods and results. I'd suggest the authors to use present tense in reporting results in the Abstract, Results and Discussion sections.

2. I did some literature search for my own manuscript suggested by a reviewer to find that "dispersal ability" is a more commonly used term than "dispersal capability" in the field of ecology and evolutionary biology. I'd suggest to replace "dispersal capability" with "dispersal ability" throughout the manuscript.

3. Please provide references for the statement in line 170.

4. Delete "very" in line 181.

5. The authors' suggestion of recent expansion and high connectivity of regional T. gigas is valid based on the results of analyses. To support the genetic analyses, authors may want to find evidences from survey data to confirm the dispersal ability of T. gigas. In the third paragraph of discussion, authors seem to list evidences on the dispersal ability of T. gigas, however the logic flow is weak. I assume the authors trying to suggest that T. gigas may disperse further based on long-term tracking in contrast to the previous suggestions of limited dispersal ability by short-term tracking studies. I'd suggest authors to add a few statement of their comments on the dispersal ability of T. gigas.
Experimental design: No comment
Validity of the findings: No comment
Additional comments: The manuscript is well written. The results and conclusion are relatively robust. After some minor changes, as suggested above, I recommend the publication of this manuscript.


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

Input:
Title: WHAT’S FOR DINNER THIS TIME?: DNA AUTHENTICATION OF “WILD MUSHROOMS” IN FOOD PRODUCTS SOLD IN THE USA
Review round: 1
Reviewer: 1

Output:
Basic reporting: - very well written

- the references are good overall although I provided a few additional references for the authors to consider. some DNA barcoding has been done for medicinal fungi and medicinal plants, which may be worth mentioning (since the sample problems were found in those studies)

- very professional and well done

- since it was descriptive there were not truly hypotheses but in this case that seemed appropriate
Experimental design: - research question was well defined and certainly this is important for food safety and for conservation

- rigorous investigation although number of reads for the next gen sequencing was low. I made some comments on the manuscript regarding the fact that it would be good to see information on negative controls and any potential contamination sources (see PDF)
Validity of the findings: - certainly this is relevant information and it has not been previously assessed in this way. it is definitely complimentary to other findings in medicinal plants, fungi, and other examples given in the intro on sushi and meat products
- underlying data seem solid and the only exception is more info needed on sequence processing and potential contamination issues (see above)
- overall very interesting work and the finding of a potentially toxic species in edible fungi packets being sold commercially is concerning and should grab the attention of regulators
Additional comments: I enjoyed this paper and think it is a solid and useful contribution. Please see detailed comments on the PDF that may help in the final revisions.


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: WHAT’S FOR DINNER THIS TIME?: DNA AUTHENTICATION OF “WILD MUSHROOMS” IN FOOD PRODUCTS SOLD IN THE USA
Review round: 1
Reviewer: 2

Output:
Basic reporting: In general well written, well explained pilot study to show the problems with the lack of labeling on mushroom products.
The strength lies in the fact that this study has been undertaken, and that even in this small sampling (only 12 products were tested), the results are surprising and scary.
The weakness is in the small sampling size, and the absence of standard sampling of the products; how many mushroom pieces were used for each product and how were they chosen? A second weakness is the use of only ITS2 for identification; the whole ITS region is considered the fungal barcode gene region, but in some groups ITS2 lacks the same level of specificity.

This is actually not the first time a study of commercial food products labled as contaning fungi is conducted; see Albertsen in  Svampe 2 (1980) on truffles in food (in Danish). [http://www.svampe.dk/svampe/SVAMPEpdf/svampe2.pdf]

The introduction lays the ground work for the issues that are investigated, the emphasis in the article should be on the USA; the EU and separate EU countries have their own legislation which is in many cases more precise than that in the USA.
It would be helpful to clearly indicate which information at the moment is mandatory in the USA - for instance, is country of origin required ? is a list of species required for imported products? and if so should they be common names or scientific names ? Are there differences between the sales of fresh wild-collected mushrooms and mushrooms that are dried or are processed (this in particular is not very clear in the introduction).
How do the regulations for herbal products differ or not from those for mushroom products?
The stress in this article is on the poisonous mushrooms, but many people experience unpleasantness after eating species that are not considered poisonous, but for which they are sensitive. More emphasis on this would be welcome.
It feels artificial to include some extra sentences on habitat degradation and decline of harvested species (in introduction and conclusions). This is beyond the scope of this article, here it is about “is what is on the lable indeed in the product?”

For readers who are not familiar with which species are cultivated and which ones are wild, it is necessary to indicate that in Table I, and also to translate the common names used on lables into scientific names, as used in the results column, in some cases that should be a genus name, such as for ‘porcini’ and ‘woodear’.

Also in Table I the contents of WMF2 are given with Latin names -common names are listed on the product, and these are in fact quite accurate (only porcini were not found in the product)

the English is good to follow, but there are a few awkward sentences and grammatical mistakes (awkward wording at lines 14–15 and 42-43; line 48, 80 - should be plural instead of singular)  Table I WMFD contains Pulchroboletus rubricitrinus.

References for the effects of Paxilus involutus and allies should be for instance the following (not a guide book!):
Flammer, R. Paxillus syndrome: immunohemolysis following repeated mushroom ingestion. Schweizerische Rundschau für Medizin Praxis 74, 997–999 (1985).
Bschor, F., Kohlmeyer, J. & Mallach, H. J. Neue Vergiftungsfälle durch Paxillus involutus. Zeitschrift für Pilzkunde 29, 1–3 (1963).
Winkelmann, M., Stangel, W., Schedel, I. & Grabensee, B. Severe hemolysis caused by antibodies against the mushroom Paxillus involutus and its therapy by plasma-exchange. Klinische Wochenschrift 64, 935-938, doi:10.1007/bf01728620 (1986).

references -
- could you check those for Gelardi and (use capital letters for genus names),and Zhao (capital letters, pages 1127–1136)
- for Von Hagen et al. i found the following: Von Hagen, B., Weigand, J. F., McLain, R., and Fight, R. (1996). Conservation and development of nontimber forest products in the Pacific Northwest: an annotated bibliography. Gen. Tech. Rep. PNW-GTR-375. Portland, OR: U.S. Department of Agriculture, Forest Service, Pacific Northwest Research Station. 246 p. DOI: 10.2737/PNW-GTR-375
- Lowenstein et al. : PLoS ONE 4(11): e7866.
- is there a web site for  “Nair, M. P. (2016). National survey of state regulation of wild mushroom foraging for retail sale. AFDO Board of Directors , page 5.”? I failed to find this.
Experimental design: see above under strengths and weaknesses
Validity of the findings: For the results - the name Suillus granulatus is used for several different species, could you specify which ones were found in the products that were sampled.
In line 155 it is stated that WMFE contains a Paxillus species - i assume WMFA is meant.
and in Figure 2 - the fragment in the upper left hand part of the picture also looks like a gilled mushroom (a Russula or an Amanita species).
Could you, based on the results of your identifications, say from which geographical area the products came from? I only saw one country of origin (Bulgaria for WMF4).
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: WHAT’S FOR DINNER THIS TIME?: DNA AUTHENTICATION OF “WILD MUSHROOMS” IN FOOD PRODUCTS SOLD IN THE USA
Review round: 2
Reviewer: 1

Output:
Basic reporting: In general my comments are minor and have to do with clearness of the text, nuancing and extra information. These are indicated in the pdf file.

I recommend structuring the methods section, and starting out saying that three different methods were used to extract and sequence the mushroom bits, followed by an elaboration of these methods.

The sample designation in table 1 is quite unclear, and could be explained more clearly in the legend. Are the replicates done with the same methods or different ones? and what doe the a and b annotations stand for?

The list of references needs your attention, as capital letters have been changed to lower case (e.g. china instead of China, and its for ITS).
Experimental design: the only comment i have is on the sampling and mixing of the samples (see the pdf); i recommend more clarity in this section.
Validity of the findings: I recommend that the authors read more about mushroom poisonings, and assumed mushroom poisonings in European mycological journals; for instance in Germany there were cases of severe poisonings by people eating Chinese dried boletes which contained parts of leaves of Araceae (perhaps used for packaging); these were the culprits for the poisonings, and not the mushrooms.

As for protection of mushrooms in the USA, not a single mushrooms species is included in the Endangered species act, but regionally, they are included in conservation efforts, e.g. in the Northwest Pacific Forest Plan. And globally a number of fungal species is now listed in the IUCN red data list, but that list is an advocacy tool. These include species that are local in the USA.
Additional comments: Thank you for incorporating the comments and requests made by the reviewers.

In general my comments are minor and have to do with clearness of the text, nuancing and extra information. These are indicated in the pdf file.

my other recommendations are in the two boxes above and in the text itself.


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: NEURAL CORRELATES OF TACTILE HARDNESS INTENSITY PERCEPTION DURING ACTIVE GRASPING
Review round: 1
Reviewer: 1

Output:
Basic reporting: The manuscript is easy to understand but needs to be sent for English proofread.

Some of the literature refences are old but overall is sufficient to the background/context of the study.

I would suggest to include;

Please include flow chart or figure with description for the experiment procedure, step by step. This will make easier for reader to understand how the experiment was conducted.

Please include figure with description for the experimental paradigm.
Experimental design: 2.	In the abstract, the is no details on the participants, data analysis and how the tactile intensities were ratings.

3.	In the method,

please also include the mean age for the female volunteers.
Twenty-five healthy volunteers (9 males, mean age: 24±4 years) participated in this study
And please also include the age range for volunteers.

Please include flow chart or figure with description for the experiment procedure, step by step. This will make easier for reader to understand how the experiment was conducted.

Please include figure with description for the experimental paradigm.

Participants were instructed to grasp the stimuli with constant force, and to focus on the hardness of the stimuli during the grasping action.
How do you control the grasp force action among the volunteers?  How to make sure that all the volunteer´s force is not varies as this will contribute to brain activation intensities.

We excluded four participants from all data analyses because of excessive head movements
I am not clear here, the total participants would be 21 or, 25 participants?
21 participants for fMRI study would be under power, and we have to consider gender differences.
Abdouni et al 10.1038/s41598-018-32724-4 reported the differences between male and female processing tactile perception.
My suggestion to add more participants to have an adequate statistical power. It is difficult to draw a conclusion with only 21 participants.
Validity of the findings: The present study only have 21 participants.

21 participants for fMRI study would be under power, and we have to consider gender differences.
Abdouni et al 10.1038/s41598-018-32724-4 reported the differences between male and female processing tactile perception.
My suggestion to add more participants to have an adequate statistical power. It is difficult to draw a conclusion with only 21 participants.
Additional comments: In this study, the authors looked at that neural responses of primary or secondary somatosensory areas that would represent the hardness intensities during active grasp exploration. They found significant BOLD responses in the posterior insula in the right hemisphere and the right posterior lobe of the cerebellum, which parametrically tracked hardness intensity. This is the first study to reveal neural correlates of the perception of hardness intensity during active tactile exploration in humans. While this is an interesting topic, the current manuscript has several limitations.

1.	The manuscript needs to be sent for English proofread.

2.	In the abstract, the is no details on the participants, data analysis and how the tactile intensities were ratings.

3.	In the method,

please also include the mean age for the female volunteers.
Twenty-five healthy volunteers (9 males, mean age: 24±4 years) participated in this study
And please also include the age range for volunteers.

Please include flow chart or figure with description for the experiment procedure, step by step. This will make easier for reader to understand how the experiment was conducted.

Please include figure with description for the experimental paradigm.

Participants were instructed to grasp the stimuli with constant force, and to focus on the hardness of the stimuli during the grasping action.
How do you control the grasp force action among the volunteers?  How to make sure that all the volunteer´s force is not varies as this will contribute to brain activation intensities.

We excluded four participants from all data analyses because of excessive head movements
I am not clear here, the total participants would be 21 or, 25 participants?
21 participants for fMRI study would be under power, and we have to consider gender differences.
Abdouni et al 10.1038/s41598-018-32724-4 reported the differences between male and female processing tactile perception.
My suggestion to add more participants to have an adequate statistical power. It is difficult to draw a conclusion with only 21 participants.

4.	In the discussion,
It is unclear, how author correlate between behavioural score and fMRI data and how this difference intensities of tactile correlate to behavioural data.
Authors did not describe the activation pattern according to the difference intensities of the tactile. Please also tabulate in the table the activation intensities vary in response to textile intensities.
Please also discuss in detail the activation areas evoked by this stimulus.

Thank you.


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

Input:
Title: NEURAL CORRELATES OF TACTILE HARDNESS INTENSITY PERCEPTION DURING ACTIVE GRASPING
Review round: 1
Reviewer: 2

Output:
Basic reporting: It is suggested to includes recent citations as references for tactile and somatosensory study in the introduction and discussion section.

Below are few recent articles that suggest being added. Please find few others in the area to support your statement in the introduction, support your findings and critically discuss it.
•	Seri FAS, Abd Hamid AI, Abdullah JM, Idris Z, and Omar H (2020). Brain responses to high frequencies (270 Hz-480 Hz) changes due to vibratory stimulation of human fingertips: An fMRI study. J. Phys.: Conf. Ser. 1497 012012. doi:10.1088/1742-6596/1497/1/012012
•	Seri FAS, Abd Hamid AI, Abdullah JM, Idris Z, and Omar H. (2019). Brain responses to frequency changes due to vibratory stimulation of human fingertips: An fMRI study. J. Phys.: Conf. Ser. 1248 012029. doi:10.1088/1742-6596/1248/1/012029
•	Vuong QC, Shaaban AM, Black C, Smith J, Nassar M, Abozied A, Degenaar P and Al-Atabany W (2020) Detection of Simulated Tactile Gratings by Electro-Static Friction Show a Dependency on Bar Width for Blind and Sighted Observers, and Preliminary Neural Correlates in Sighted Observers. Front. Neurosci. 14:548030. doi: 10.3389/fnins.2020.548030
•	Kim Y, Usui N, Miyazaki A, Haji T, Matsumoto K, Taira M, Nakamura K and Katsuyama N (2019) Cortical Regions Encoding Hardness Perception Modulated by Visual Information Identified by Functional Magnetic Resonance Imaging With Multivoxel Pattern Analysis. Front. Syst. Neurosci. 13:52. doi: 10.3389/fnsys.2019.00052

It is suggested that any tactile and somatosensory studies used other brain imaging modalities like EEG, MEG, etc., to be included in the discussion section. This will help to discuss some of your findings and limitation.

Example study in the related areas but using EEG.
•	Baghdadi, Golnaz, Amiri, Mahmood, Falotico, Egidio, Laschi, Cecilia. 2020. Recurrence quantification analysis of EEG signals for tactile roughness discrimination. International Journal of Machine Learning and Cybernetics. https://doi.org/10.1007/s13042-020-01224-1

Please move the limitation in the conclusion section to the discussion part. Authors can put it under limitation and future suggestion subheading.
Experimental design: Data analysis:
1.	Please include the information of acceptable head movement to be included as the participants of this study.
2.	Please include the design matrix used in the first level analysis/GLM
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: NEURAL CORRELATES OF TACTILE HARDNESS INTENSITY PERCEPTION DURING ACTIVE GRASPING
Review round: 2
Reviewer: 1

Output:
Basic reporting: The author(s) had done the amendments accordingly, and I have no further suggestions.
Thank you.
Experimental design: The author(s) had done the amendments accordingly, and I have no further suggestions.
Thank you.
Validity of the findings: The author(s) had done the amendments accordingly, and I have no further suggestions.
Thank you.
Additional comments: The author(s) had done the amendments accordingly, and I have no further suggestions.
Thank you.


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

Input:
Title: NEURAL CORRELATES OF TACTILE HARDNESS INTENSITY PERCEPTION DURING ACTIVE GRASPING
Review round: 2
Reviewer: 2

Output:
Basic reporting: The revised version is clear and well structured.
No comment
Experimental design: No comment
Validity of the findings: No comment
Additional comments: Overall, the authors address all questions raised by the reviewers clearer and well structured. The authors also provided few figures to better explain in visual form the paradigm and task used for this 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: LONGITUDINAL VARIATIONS IN THE GASTROINTESTINAL MICROBIOME OF THE WHITE SHRIMP, LITOPENAEUS VANNAMEI
Review round: 1
Reviewer: 1

Output:
Basic reporting: The manuscript does a reasonably good job at describing the microbial communities in white shrimp but it significantly lacks adequate interpretation of the results. This is especially evident in the underannotated figures (i.e. what do the values mean in Fig. 2; what are the values in the heatmap in Fig. 6). Additionally, there's several minor grammatical errors that make comprehending the narrative difficult.
Experimental design: The manuscript fails to adequately describe the experimental design (i.e. L107-L118, how many samples were included in the analyses; were the samples comprised of pools of shrimp gut tissues or individual tissues), making it difficult to understand how well the observed results can be generalized across white shrimp. This should be made much clearer.
Validity of the findings: The findings, while lacking in interpretation, seem fine, but this is somewhat hard to assess given that it is unclear what the dataset looks like.
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: LONGITUDINAL VARIATIONS IN THE GASTROINTESTINAL MICROBIOME OF THE WHITE SHRIMP, LITOPENAEUS VANNAMEI
Review round: 1
Reviewer: 2

Output:
Basic reporting: The paper entitled: “Microbiota variations through the gastrointestinal tract of the white shrimp, Litopenaeus vannamei” by Garibay-Valdez et al, reports the microbiome composition of three portions of the GI tract in the white shrimp.
I think the study is potentially interesting, but needs improving before publication. There are some minor errors in the English language that need to be corrected (there is an abuse of the possessive case for instance). Moreover, some points of the paper are unclear to me.
I think the aim of the study should be better explained: why is it so important to look at the 3 portions, what is the “catch”. You report the economic value of the shrimp, but is the different localization of the bacterial species changing anything in the strategy of aquaculture for example? Or is it just interesting for the scientific point of view? If so you should mention.
I think the results is missing a summary of the composition of the samples, at different taxa levels, what are the main phyla, classes…etc? can you represent this in a barplot? I think it would be nice to see it graphically to highlight any difference among the 3 sampling sites.
In general, I think some parts of the manuscript would be better presented in a table, ie the Pathways in line 300-331. I see you refer to tab1 but there is no mention in the tables that these Pathways are the one overrepresented.
Experimental design: First of all, how did you sample the 3 portions of the GI? Did you sample them from the same animals? I think the best way to account for individual variability would be to sample the 3 portions from all the animals. You did not justify why you have a different number of samples for the 3 portions. I could not see any reference to the final number of samples the authors studied. Moreover, why did you divide in groups? How many animals/group? What are the differences among the groups?
Finally. The results of alpha and beta-diversity need to be improved: is there a difference in observed ASV number and/or Shannon index in the 3 portion of the gut? What are the R2 and P values for the 3 distances you calculated?
In general, why did you decide to focus on the families only? What about at higher taxonomic levels?
Did you sample the water? It would be great to have that info to see if some of the species are found there.
Validity of the findings: I cannot comment on this before I understand how many samples where included in the study.
I think the novelty of the study is well stated, but I do not get the importance of studying the difference localization of the bacterial community.
Additional comments: please refer to my previous comments to implement the paper. Find here also some more detailed comments:

Title: I do suggest to use “Longitudinal variations of the microbiome of the gastrointestinal tract of the white shrimp, Litopenaeus vannamei”
Line 28: of the foregut
Line 30 and 37: I would not use “potential”
Line 42-43: relies on the fraction… is not clear, please rephrase.
Line 44-45: providing… is not clear, please explain.
Line 49-50: I would report here the main diseases that interest this species.
Line 58: ‘Provides beneficial host functions…” is confusing. Please rephrase
Line 59-62: this sentence is missing the verb
Line 70: maybe use “digestive”
Line 75: this sentence is confusing
Line 98: at this point the authors should state the number of animals
Line 107: what are the differences among the groups?
Line 107-8: I am confused on the number of shrimps: 5 groups *3 replicates*20 animals?
Line 111: use always the same units (you used before mg/Ml at lune 101, then you used the “-1’.
Line 113-114: what is “almost 0”?
Line 131: please report the reference for the primers
Line 140: past of load is loaded
Line 148: denoised
Line 165: did you calculate any alpha-diversity indexes?
Line 179: are these the total numbers of
Line 180 and 215: I would use clustered or parsed?
Fig1: I am not sure what the graph represents
Fig3 is cut in two points
Line 235: it is ASVs not ASV’s
Fig4: there is a typo in the caption. How many animals you had? How you did not have the same number for the 3 locations? Did you extract the DNA from the same samples? If you did not, how can you be sure the differences are not due to the individual variability?
Line 245: can you provide the results of the beta-diversity tests? Including R2 and P values
Line 257: use the past tense and the same all along the manuscript
Line 278: is this order in the “taxonomic group” sense?
The figures are not in the right order.
Line 301-2: is this true for the 3 locations?
Line 308: fractions
Fig7 can go on the supplementary material
Why didn’t you put a Ven diagram to report the shared taxa? And why did you chose to report only family level?
Line 433: and in italics


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: LONGITUDINAL VARIATIONS IN THE GASTROINTESTINAL MICROBIOME OF THE WHITE SHRIMP, LITOPENAEUS VANNAMEI
Review round: 1
Reviewer: 3

Output:
Basic reporting: .
Experimental design: .
Validity of the findings: .
Additional comments: I’ve read and analized the work carefully.
It is an original work, in which various tools where used, including molecular biology like bioinformatics to breakdown since the holobionte point of view to the white shrimp Litopenaeus vannamei. Comparing the three most important components in the digestic tract: the foregut (stomach), midgut (hepatopancreas), and hindgut (intestine). With the goal of knowing the genetic structure of the bacterial community using 165 rRNA.
To contribute in the understanding of the functions of said bacterial communities.
The manuscript is clear, professional English language used throughout. I just have minor suggestions for page 6 line 38 it says while, I suggest while.
Lines 40 repeats “and” twice.
Line 257 says “vary” I suggest “varies”
Lines 484 also repeats “and” twice.
The results are fresh, new, and they contribute to the knowledge of the microbiota paper. The structure is clear, the Figures are relevant, well labelled & described.
Rigorous investigation performed to a high technical.
The results are clearly described, for disscusion there is an extensive bibliographic revision which is actualized and opportune.
All of the quotes are on the literature and the other way around.


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: LONGITUDINAL VARIATIONS IN THE GASTROINTESTINAL MICROBIOME OF THE WHITE SHRIMP, LITOPENAEUS VANNAMEI
Review round: 2
Reviewer: 1

Output:
Basic reporting: The 'new' figure legend for figure 6 does not correspond with what is presented in the figure.
Otherwise, the revisions made, especially those highlighted by the co-reviewer, improve the manuscript.
Experimental design: sufficiently improved.
Validity of the findings: Reasonable.
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: LONGITUDINAL VARIATIONS IN THE GASTROINTESTINAL MICROBIOME OF THE WHITE SHRIMP, LITOPENAEUS VANNAMEI
Review round: 2
Reviewer: 2

Output:
Basic reporting: I was very happy to revise the paper again and notice how it improved after the first round of revisions.
Minor comments:

Unit are still written in different ways: mg/ml or ml-1 or day-1, also check ml or mL
Line 117: shrimps
Line 144: we only kept
Line 195   and 275: 4,999
Line 243: fig s2 csn go to main text
Fig 3 is still cut in some points: ie close to rubritaleaceae
Experimental design: the experimental design is now fully explained in the revised paper.
Validity of the findings: no comment
Additional comments: I very much appreciated the effort the authors put in the revision.


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: DEVELOPMENTAL INSTABILITY IN WILD NIGERIAN OLIVE BABOONS (PAPIO ANUBIS)
Review round: 1
Reviewer: 1

Output:
Basic reporting: The article is written in a clear and direct manner that avoids the overuse of jargon.  A concise overview of fluctuating asymmetry is provided, along with a brief discussion of factors that may confound such an analysis.  Appropriate references are made to previous research on the subject (specifically, Palmer and Strobeck, Bailit, Garn, Perzigian, etc...).  The structure and organization of the article is good; I found little reason to backtrack, as each section builds in an appropriate way on the last.  Figures and tables are labeled appropriately and convey the results of the research well.

There were, however, several things that I felt might have improved the communication of this research to an audience.  It would have been helpful for the authors to include the actual ages (in years or months) at which weaning and sexual maturity occur in relation to when crown development is completed, as a means of demonstrating that the timing of these stressors is correlated to crown asymmetry.  It also might have been useful to provide additional comments within the R scripts in the data repository or file names that communicate the purpose/function of these files -- the data and the scripts themselves, however, were excellent.  It might also have been beneficial to include a table of the ANOVA results.

There is a typo on line 82 -- I believe this is meant to read "is non-directional" rather than "in non-directional"
Experimental design: The research, which tests three interrelated hypotheses regarding fluctuating asymmetry (FA) in baboons, certainly fits within the aims and scopes of the journal.  The research questions are well defined and the purpose of the research itself (as FA has been largely ignored in non-human primates) is certainly relevant.

The experimental design follows the recommendations of Palmer and Strobeck (1986) and Palmer (1994) for an analysis of fluctuating asymmetry.  Measurement error was estimated for replicate data, outliers were removed when appropriate, and the resulting data sets were tested for deviations from normality and confounding factors such as directional asymmetry and antisymmetry.  Estimation of FA, utilizing the FA10a index value which removes measurement error as part of a two-way mixed model ANOVA, resulted in valid estimates of FA for the groups under study.

I felt that two aspects of the methods might have been discussed in more detail.  The actual process of determining whether or not these traits were size dependent (and why this is important) might have been discussed in more detail.  Likewise, the method for detecting antisymmetry through an analysis of skewness and kurtosis could also be outlined in more detail.  The article, on line 233-235 suggests that these methods are outlined in the data repository -- rather than go through an external source, I would have liked to see this discussion in the article itself.
Validity of the findings: The test results for the three hypotheses are clear and direct; the authors demonstrate through their analysis that estimates of FA differences between males and females, and differences by sex with regards to weaning and sexual maturity, were significant.  The authors have, generally, avoided unnecessary speculation about the cause and effect relationship between FA and specific stressors.

I do question, however, the reference to Bailit (1970) on line 330.  The authors suggest that human studies of FA are inappropriate for comparison to non-human primates (lines 105-111) yet they are comparing their own results to those of Bailit which, in addition to being based on a human sample, are based upon a less rigorous analysis of FA.
Additional comments: A well designed and executed analysis of fluctuating asymmetry within Papio anubis that makes a long overdue and meaningful contribution to the scholarship on developmental stress in non-human primates.


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: DEVELOPMENTAL INSTABILITY IN WILD NIGERIAN OLIVE BABOONS (PAPIO ANUBIS)
Review round: 1
Reviewer: 2

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: This is a very nice contribution to the science of non-human primate dental studies, with relevance to mammalian growth and development more generally. The authors have written a clean manuscript that makes their study design clear, their results easy to see, and their conclusions in line with the scope of the strengths of the study design and their results. The observation that FA varies differentially between males and females is very interesting. This window into the stress differences that males and females experience during their maturation is intriguing.

I have just three recommendations:
1)	Add a sentence to the abstract noting the environmental “extreme-ness” of this population among baboons.
2)	Add in a few sentences answering the question of how interstitial wear may be playing into the FA magnitude difference observed between mesiodistal and buccolingual measurements.
3)	Make the colors and labeling of the three bar charts more user-friendly.


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: DEVELOPMENTAL INSTABILITY IN WILD NIGERIAN OLIVE BABOONS (PAPIO ANUBIS)
Review round: 1
Reviewer: 3

Output:
Basic reporting: The language use is standard and clear. I have noted a few minor exceptions with comments in the marked up pdf of the manuscript.

The literature is appropriate, though I feel the authors missed an opportunity to strengthen their argument for the use of FA as a stress indicator, by not citing any of the literature that addresses problems with identification and error associated with the analysis of dental defects.

The structure, tables, and figures are appropriate, and the authors state they are sharing their raw data.

The hypotheses are testable and tested. I suggest the rewording of one hypothesis, and the possible re-consideration of a second. This reconsideration should not be a requirement of publication.
Experimental design: We know almost nothing about NHP dental FA, so this paper definitely fills a gap.

The investigation is rigorous, although perhaps more than need be. I don't understand why the authors took 10 sets of measures. An explanation for this choice should be made, especially as they point out that it made their analysis more complicated.

The methods are duplicable as described.
Validity of the findings: The only issue here is the overstatement of the purity of results. FA is a result of interactions among environmental stressors and somatic buffering/frailty. The authors do not weigh their results interpretations sufficiently because they do not address the unknown effects of buffering. Please see specific comments made in the MS pdf.

Otherwise, all is reasonable.
Additional comments: Thank you for writing a straightforward, easy to read paper, that makes a very specific contribution. If you address the comments made here and in the pdf, the paper should be publishable in short order.


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: RNASE H-DEPENDENT AMPLIFICATION IMPROVES THE ACCURACY OF ROLLING CIRCLE AMPLIFICATION COMBINED WITH LOOP-MEDIATED ISOTHERMAL AMPLIFICATION (RCA-LAMP)
Review round: 1
Reviewer: 1

Output:
Basic reporting: The manuscript has a clear and professional use of English; some suggestions to improve some sentences are given in the word file. The references used in the manuscript are the adequate to show how accuracy of a method can be obtained. The paper´s structure is good, and figures are clear and concise.
In line 189 of the word file, it is unclear who or what "This" refers to, could you please consider rewriting the sentence to make it clearer?
Experimental design: The methods are clearly described, the DNA and RNA sequences designed and used are clearly presented. Conditions for the amplification reactions are detailed and clearly presented.
Validity of the findings: An improved RCA-LAMP technique with improved accuracy is presented in the manuscript with the introduction of RNase H-dependent PCR. This technology is desirable specially when amplifying very small amount of RNAs, like small RNAs.

I consider this improvement important, since non-specific amplification is not desirable in PCR reactions. I think the data presented is robust and conclusion is clearly stated and supported by the data.
Additional comments: The manuscript presents in a clear way how to improve the accuracy of the RCA-LAMP isothermal amplification to quantify RNA. For the real-time quantification did you did any statistics? I did not see at the manuscript the number of technical replicates for the experiments.


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: RNASE H-DEPENDENT AMPLIFICATION IMPROVES THE ACCURACY OF ROLLING CIRCLE AMPLIFICATION COMBINED WITH LOOP-MEDIATED ISOTHERMAL AMPLIFICATION (RCA-LAMP)
Review round: 1
Reviewer: 2

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The authors present an interesting and straightforward set of experiments. They focused on the technique of RNase H-dependent PCR (rhPCR). By applying this technique, the accuracy of RCA-LAMP is improved. They found rh primer and RNase H2 enzyme suppressed the non-specific amplification of linear PLP. Further, non-specific amplification in a traditional RCA-LAMP assay was discussed. As such, the work presented in this manuscript provide useful information for researchers in this field.


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: RNASE H-DEPENDENT AMPLIFICATION IMPROVES THE ACCURACY OF ROLLING CIRCLE AMPLIFICATION COMBINED WITH LOOP-MEDIATED ISOTHERMAL AMPLIFICATION (RCA-LAMP)
Review round: 1
Reviewer: 3

Output:
Basic reporting: In this manuscript, the authors proposed the method for suppressing the non-specific amplification signal in RCA-LAMP technology, which relies on RNase H-dependent PCR primer reported in 2011 (ref 1). The proposed idea is worthy of publication, but there are many missing and incorrect results in the manuscript. Therefore, I suggest the acceptance of this manuscript after accommodating the following comments.

Ref 1) BMC Biotechnology, 2011, 11: 80
Experimental design: 1) The title should be changed because it sounds like PCR is used in this work.
2) In Figure 3 & 4, the authors did not test the forward primer with RNA modification only (rh (-,+)). The authors need to provide the results obtained by RCA-LAMP with rh (-,+).
3) The authors added RNA at the 6th from the 3’ end of primer. In the original rHPCR paper (ref 1), RNA is added at the 5th from the 3’ end of primer. The authors need to provide the reason for this or optimization results.
Validity of the findings: 1) In Figure 4, the authors need to provide more details about how to calculate POI
2) In Figure 4(D), xrh (-,+) should be corrected into xrh (+,-). In addition, Figure 4(A-C) do not match with the results in Figure 4(D), and the line color in Figure 4(A-C) is unclear. The authors need to change these mistakes.
3) The authors need to calculate the limit of detection, and test the target RNA at lower concentrations (e.g. 10 fM, 1 fM, 100 aM). In the original RCA-LAMP paper (ref 2; Biosensors and Bioelectronics, 2019, 128: 17-22) achieved the limit of detection of 10 aM without rHPCR technique, which seems better than the one in this work.
4) The authors emphasized that the use of RNA incorporated primer can suppress the non-specific amplification signal produced in the absence of target RNA, but undesirable amplification in the absence of RNA was observed in Figure 4. The authors need to discuss why these background signals do not completely disappear.
5) The authors need to check the specificity of the proposed system by using the control RNA samples.
6) Instead of listing the primer sequences, the authors need to make the table for clarity.
Additional comments: In this manuscript, the authors proposed the method for suppressing the non-specific amplification signal in RCA-LAMP technology, which relies on RNase H-dependent PCR primer reported in 2011 (ref 1). The proposed idea is worthy of publication, but there are many missing and incorrect results in the manuscript. Therefore, I suggest the acceptance of this manuscript after accommodating the following comments.

Ref 1) BMC Biotechnology, 2011, 11: 80
Ref 2) Biosensors and Bioelectronics, 2019, 128: 17-22

1)	The title should be changed because it sounds like PCR is used in this work.
2)	In Figure 3 & 4, the authors did not test the forward primer with RNA modification only (rh (-,+)). The authors need to provide the results obtained by RCA-LAMP with rh (-,+).
3)	The authors added RNA at the 6th from the 3’ end of primer. In the original rHPCR paper (ref 1), RNA is added at the 5th from the 3’ end of primer. The authors need to provide the reason for this or optimization results.
4)	In Figure 4, the authors need to provide more details about how to calculate POI
5)	In Figure 4(D), xrh (-,+) should be corrected into xrh (+,-). In addition, Figure 4(A-C) do not match with the results in Figure 4(D), and the line color in Figure 4(A-C) is unclear. The authors need to change these mistakes.
6)	The authors need to calculate the limit of detection, and test the target RNA at lower concentrations (e.g. 10 fM, 1 fM, 100 aM). In the original RCA-LAMP paper (ref 2) achieved the limit of detection of 10 aM without rHPCR technique, which seems better than the one in this work.
7)	The authors emphasized that the use of RNA incorporated primer can suppress the non-specific amplification signal produced in the absence of target RNA, but undesirable amplification in the absence of RNA was observed in Figure 4. The authors need to discuss why these background signals do not completely disappear.
8)	The authors need to check the specificity of the proposed system by using the control RNA samples.
9)	Instead of listing the primer sequences, the authors need to make the table for clarity.


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: RNASE H-DEPENDENT AMPLIFICATION IMPROVES THE ACCURACY OF ROLLING CIRCLE AMPLIFICATION COMBINED WITH LOOP-MEDIATED ISOTHERMAL AMPLIFICATION (RCA-LAMP)
Review round: 2
Reviewer: 1

Output:
Basic reporting: The reviewed version of the manuscript is clearly presented. Authors answered each of the reviewers´ questions in a proper way. Title is now more appropriated and professional english was used throughout the manuscript. Figures are well presented, and a table was added to show the primers´ sequences utilized in experiments. The paper is now fluid and readable. The paper structure is appropriated for Peer J.
Experimental design: All questions raised by reviewers are answered and proper reasons are given at each question raised. A legend about number of replicates was added to Figure 4. I consider this part is well described in the manuscript in a clear way.
Validity of the findings: Points to review and correct were tackled and now it is in a good shape. Authors improved Results and Discussion sections taking into account points raised during the first revision. Conclusion is clearly stated.
Additional comments: The paper "RNase H-dependent amplification improves the accuracy of rolling circle amplification combined with loop-mediated isothermal amplification (RCA-LAMP)" presents experimental evidence of the improvement of accuracy of the RCA-LAMP technique. The work is clearly presented, and the figures are precise.


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: BRAIN DERIVED NEUROTROPHIC FACTOR DECLINES AFTER COMPLETE CURATIVE RESECTION IN GASTROINTESTINAL CANCER
Review round: 1
Reviewer: 1

Output:
Basic reporting: The manuscript is clearly written in professional, unambiguous language.
Experimental design: This is a well-designed study that systematically and robustly addressed a scientific hypothesis.
Validity of the findings: no comment
Additional comments: 1. The article draws a conclusion that our results suggested that BDNF might play an important role in gastrointestinal cancer development. BDNF levels were correlated with tumour volume, and with neuro-, angio-, and lymphatic invasions. It may have a few deficiencies. If the author can include the serum BDNF concentration of normal people, and compare the serum BDNF concentration between the cancer patients and the normal people, this will make the article more convincing.
2. Line 78-91: more details about the role of BDNF in angiogenesis, proliferation, cell migration, and apoptosis are needed.
3. Line 90-91: There may be relevant studies about the role of serum BDNF concentration in cancer (DOI: 10.1155/2021/8867368), the author can refer to related research.
4. Line 120: Why did the author choose to detect the serum concentration on the third day after operation. Is longer time reasonable.
5. In the results section, the author mainly presents it in the form of tables. If it can be in the form of histogram or other forms, it will make the article better present the consequences.
6. All tables in the article should be a three-line table.
7. Line 287-289: Maybe the article has more limitations.
8. If the author can detect the expression of BDNF in tumor, this will make the article more convincing.


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: BRAIN DERIVED NEUROTROPHIC FACTOR DECLINES AFTER COMPLETE CURATIVE RESECTION IN GASTROINTESTINAL CANCER
Review round: 1
Reviewer: 2

Output:
Basic reporting: This manuscript is a very timely look at the serum levels of BDNf in patients with colorectal cancer. The text is clear and easy to read and comprehend. There is enough background and context provided.
Experimental design: The experimental design is suitable for the topic and type of study and the methods are easy to follow.
Validity of the findings: The findings are valid  with robust underlying data provided. The conclusions are good and not over-reaching, given the small dataset.
Additional comments: An interesting and well carried out small study as a snapshot of BDNf in this patient cohort.


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: BRAIN DERIVED NEUROTROPHIC FACTOR DECLINES AFTER COMPLETE CURATIVE RESECTION IN GASTROINTESTINAL CANCER
Review round: 1
Reviewer: 3

Output:
Basic reporting: The paper is adequately prepared.
Experimental design: Design, procedures and analyses are adequate.
Validity of the findings: Although it is not a large study, the number of subjects is sufficient to provide clear results.
Additional comments: Although BDNF/TrkB signaling have been increasingly investigated in cancer, the role of serum BDNF and its potential role as a biomarker in oncology is much less explored. Thus this study comparing serum BDNF levels before and after surgery in pacreatic and colon cancer will contribute to the community working on this theme and help elaborating new hypotheses for studies of BDNF as a biomarker in cancer. The number of patients could be larger but is adequate for an early study such as this one. The paper is well written and methods, result presentation data interpretation and ethical aspects are adequate.
- Lines 201 - 207, note that serum BDNF levels have been recently also measured in pediatric leukemia and low levels were found to be associated with active disease and poor prognosis, whereas normal BDNF levels were seen when the treatement was sucessful and associated wth better prognosis (Portich et al., Low brain-derived neurotrophic factor levels are associated with active disease and poor prognosis in childhood acute leukemia. Cancer Biomark. 2016 Sep 26;17(3):347-352. doi: 10.3233/CBM-160646). Can the authors speculate on how to reconcile these findings with their own findings and other published studies?
- Also note that higher tumor gene expression (transcript levels) of BDNG and NTRK2 genes have been recently associated with lower overal survival in a solid tumor type (medulloblastoma) (Thomaz et al., Neurotrophin signaling in medulloblastoma. Cancers (Basel). 2020 Sep 7;12(9):2542. doi: 10.3390/cancers12092542).
- Finally, it is important that, in the Discussion, the authors highlight the important recentl advances in the clinical use of Trk inhibition in patients with differente types of tumors harboring NTRK fusions.


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: BRAIN DERIVED NEUROTROPHIC FACTOR DECLINES AFTER COMPLETE CURATIVE RESECTION IN GASTROINTESTINAL CANCER
Review round: 2
Reviewer: 1

Output:
Basic reporting: Authors made all requested minor revisions.
Experimental design: No comment
Validity of the findings: The results suggested that  BDNF levels were correlated with tumour volume, and with neuro-, angio-, and lymphatic invasions.Thus, BDNF may serve as a potential marker of complete resections in underdiagnosed patients.
Additional comments: Authors made all requested minor revisions.


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: VALIDITY AND USEFULNESS OF THE STUDENT-ATHLETES’ MOTIVATION TOWARD SPORT AND ACADEMICS QUESTIONNAIRE: A BAYESIAN MULTILEVEL APPROACH
Review round: 1
Reviewer: 1

Output:
Basic reporting: The manuscript presents the validation process of the SAM-SAQ/PT. The manuscript is well written and structured. The sources of information are updated and relevant.
The methodology and statistical analyses are its main strengths. The initial large sample allows the authors to conduct the validation, however, reusing with the same sample for further analyses maybe not necessary as the core of the contribution is the validation of the SAM-SAQ/PT tool.
Still, the content of this article would be a great contribution to the Dual Career topic, in case it is published.
I have made some specific suggestions which I hope the authors find them in a constructive way.
My recommendation for this manuscript is a major revision.
Experimental design: These comments are in the "specific comments".
Validity of the findings: These comments are in the "specific comments".
Additional comments: SPECIFIC COMMENTS
ABSTRACT
Review the content of the abstract based on the specific comments about the four studies.

L 25-26 & 28 Authors repeated the content " Four studies were performed."
L 31-32 I do not understand study 3
L 39 Predictions of scores ¿? I thought It was a comparison

INTRODUCTION
L 60 Authors introduce here for the first time the dual-career pathways, but this concept is not explained.
L 81-84 Please provide data about public-private universities. Which is the proportion of them in the whole country? and what is the number of students in each one?
L 84 & L 88 These references are not included in the final list. Please provide them.
L 92, 95 & 96 Please revise the references.
 (Open Science Collaboration 2015) needs a comma.
(Amrhein & Greenland 2018, needs a comma
McShane, Gal et al. 2019) should be McShane et al., 2019)
AIMS
L114-119 Please writ-state each aim (one per study) separately in this paragraph rather than "addressing".
L116-117 the SAM-SAQ IT is supported by the reference (Guidotti & Capranica, 2013; but the "A" is not included in that reference. Please revise the whole manuscript.

METHODS
I would recommend authors changing this section structure.
I had problems understanding the section itself.
First, the information is repeated or the content is not in an adequate place. For example:
	• L 131-133 there are some variables mentioned that should be included in the tools subsection.
The research design could include the ethics committee information and the inclusion criteria of the different samples of the study.
Then in each study authors could follow the structure of:
Participants- Tools-Procedure-Data analysis- Results-Discussion

L 156-157 No supplemental files are available for review.
L 157- could you give more information about the sample? How many student-athletes compete in those championships? What was the response rate? Were they representative of their population? Were they from all the regions of Brazil? What was their race? What kind of studies they were enrolled in? STEM (Sciences Technology Engineer and Mathematics), Social Sciences, Humanities, Life Sciences…

DISCUSSION study 1
I would suggest authors begin each discussion with the aim of the study and the main results found.
L 180-189 Authors should focus on the aim of this study. These lines explain each factor which could be explained in the introduction section. Authors should compare their results (3-factor solution) with the Italian, European, and Asian validation processes.
L 189-190 This speculation should be based on the results. I cannot find the link with the Exploratory Factor Analysis results.

STUDY 2
Line 222 Please explain from the 17 final items which are from SM, AM, and CM.
L 225 Supplementary Table 2 is not available to revise.
L 237-238. Could you please detail the main differences found between this SAMSAQ-PT and the other languages versions?

STUDY 3
The aim of this study is to discriminate (separe) or to compare by gender, type of university, sport level…?
Later in the paper at the study 3 results (L 282) authors state the differences found by gender….
L 243-244- Please write the independent variables as gender, sport level, type of university and athlete status.
Please explain clear the athlete status meaning. In the tools section not in data analysis.
L 247-249 In study 1 and 2 authors validated the SAMSAQ/PT with this sample. The reuse of the sample for other purposes is not justified.
L 281-284 Please divide the differences found for each factor. With this writing, it is difficult to understand the message.
For example in Figure 1 at Sport Motivation female had higher z-score levels than male, while in the text they state just the opposite. Please revise each factor and the differences found.
Another option and based on the discussion structure would be to divide this text into gender differences- sport level differences-type of university differences and student-athlete status differences. But clearly in the text, please do not mix both writings (factors with independent variables).
L 284 Supplementary Table 3 is not available to verify its content.
Why using bayesian models when you only want to compare by one independent variable?
L 289 please begin each discussion with the aim of the study and the main results.
Structure this discussion following the same structure as the results.
L 292- 294 Please revise the English grammar ("their sport one"¿?)
L 295 Please omit writing identity when measuring motivation.
Please revise the last name of these references:
L 319- Cecic Erpic and L 331 López de Subijana

STUDY FOUR
L 340- I do not agree with the authors when they propose a repeated measured design when they measured twice. Repeated measures is a concept applied to more than 2 measures.
L 351-354- I have the same concerns about re-using the sample for validating and for other studies in the same article.
L 357- Please check the writing (mon-representative).
L 386-387- I do not see these results in the Figure. They are supporting later the discussion and they are not in the Figure.
L 387 please remark clearly that the individual is gender while academic context is the type of university. They seem new variables in the text.
Discussion
Please structure the discussion as mentioned before. First, relate the results with the aim of the study.

L 396-397 Please limit the statement to the results of this study. The decrease of SM was not reported in the results section.
L 399-402 I could not appreciate this result in Figure 4 of this study. Please limit the content to the results found.

GENERAL CONCLUSIONS

L 349-340. This sentence when this theoretical framework (Rudolph et al., 2019) is not included in the introduction does not increase
the manuscript contribution.
L 440-441- I suggest authors include the data repository in….
L 444-446 This recommendation is for fostering a dual career in public universities?
L 457- I do not understand why the authors acknowledge chronological age as a limitation when it was informed in line 154.

I think authors should limit their manuscript to the main contribution: validating the SAM-SAMQ/PT.
Reusing the sample for other purposes I think is not appropriate.


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: VALIDITY AND USEFULNESS OF THE STUDENT-ATHLETES’ MOTIVATION TOWARD SPORT AND ACADEMICS QUESTIONNAIRE: A BAYESIAN MULTILEVEL APPROACH
Review round: 1
Reviewer: 2

Output:
Basic reporting: The authors present a manuscript of relevant interest. The subject falls within the scope of the journal. However, the text still needs some amendments, clarifications, and corrections.
Experimental design: Are there really four studies, or is it a single study with four phases?
Validity of the findings: no comment
Additional comments: (1) Avoid unnecessary repetition of words and phrases. Examples:
(1.1) In the abstract: "The purpose of this research was to examine the validity and usefulness of the student-athletes`motivation toward sport and academics questionnaire (SAMSAQ-PT) in Brazilian student athletes".
(1.2) In the abstract: "Four studies were performed" is written twice.

(2) Are there really four studies, or is it a single study with four phases?

(3) Lines 98 and 431, change dierent to different.

(4) Line 154: "age 21.7 ± 3.4 yrs". The notation “mean (SD)” should be used throughout instead of “mean plus/minus SD,” since this is not a confidence interval. In other words, the SD should be indicated in parentheses.

(5) Line 163. Is a total of 20000 samples sufficient for the convergence of the MCMC algorithm? How is this verified?

(6) Line 164. What is default identification restriction? A brief explanation of its meaning and usefulness should be inserted into the article.

(7) Line 165: "The minimum posterior means for a factor loading to be considered as a benchmark was set as 3". Is this a usual choice? Are there bibliographic references for this?

(8) Line 171: "Due to low factor loadings, items 11 (2.70), 18 (2.09), 25 (2.44), and 30 (1.78) were excluded". What value determines a low factor loading?

(9) Line 206: "Two chains for 10,000 iterations with 2,000 burn-in iterations and the default Stan were run". What is Stan?

(10) Line 207: "The model estimations were regularized using normal prior (0, 10) for the manifest variable (intercept) and normal prior (0, 1) for the latent variable were set". What motivated the choice of these hyperparameters? Are they non-informative priori distributions? Are these recommendations from the literature?

(11) Line 253: "...Brazilian student-athletes when grouped by gender (e.g., female and male)...". Sex and gender are different concepts, sex refers to biological characteristics and gender refers to the individual's and society's perceptions of sexuality. Do the authors believe that this differential is important in the context of this paper?

(12) Lines 269 and 380: "Two chains for 4,000 iterations with a warm-up length of 1,000 iterations to ensure convergence of the Markov chains were run". Are the authors sure that 4000 MCMC samples are sufficient for chain convergence? It seems very little to me. As a rule of thumb, we can observe if the MC error is less than 1% to 5% of the corresponding posterior standard deviation.

(13) What is the motivation for using 67% and 90% credible intervals?

(14) Line 269: "Two chains for 4,000 iterations...". Line 379: "Four chains for 4,000 iterations...". Why did one Bayesian model use 2 chains and the other used 4 chains?

(15) Was the Gelman-Rubin convergence diagnostic 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: VALIDITY AND USEFULNESS OF THE STUDENT-ATHLETES’ MOTIVATION TOWARD SPORT AND ACADEMICS QUESTIONNAIRE: A BAYESIAN MULTILEVEL APPROACH
Review round: 1
Reviewer: 3

Output:
Basic reporting: Comment on language and grammar Issues.
•	Line 98 “di*erent” “different”
•	Line 131 “analyses” “analysis”
•	Line 153 “dataset” “data set”
•	Line 357 “mon-representative” “non-representative”
•	Line431 “di*erent” “different”
Experimental design: •	Consider an even number (0.2) or (2) as the variant rather than (.05) which appears to affect some of the accuracy in each of the four models.

•	The authors Discussion section provide accurate next steps for further research.
Validity of the findings: No criticism. Great job on a difficult topic to quantify.
Additional comments: •	Good job of introducing the questionnaire in the initial 4 lines of the background section.
•	The methods are clearly defined
•	Results are clearly defined
•	Dual Career is clearly defined in lines 51,52,53, 54
•	Categories pertaining to dual career are specified: Motivation, identity, health, lifestyle, and well-being.
•	The historical acknowledgement of the SAMSAQ-PT provides a front loading to the information being introduced and analyzed.
•	Literature references are sufficient and provide support for the research. Both the findings and what needs are necessary for improvements.
•	Figures and analysis table are clearly explained and comprehensible
•	Provides evidence of Portugese’s version of the SAMSAQ-IT/A
•	Demonstrates the significance and importance of the research into increased support for Dual Career athletes.
•	Great mastery of writing in the English language. Masterful use of writing conventions


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: VALIDITY AND USEFULNESS OF THE STUDENT-ATHLETES’ MOTIVATION TOWARD SPORT AND ACADEMICS QUESTIONNAIRE: A BAYESIAN MULTILEVEL APPROACH
Review round: 2
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: Dear authors,
I find the manuscript better structured now. I would like to thank them for the effort done at this reviewing round.

- Still there are some minor issues in relation to typing errors at some references. Please check the following references in lines: 99, 102, 103, 110, 113, 118, 346, 347, 385,
- L 161- About the supplementary tables I would recommend authors to include this link each time the refer to supplementary materials as It won’t be available each content in the article.
- L 536-537 Please mention again that the supplementary tables are in that link.