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

Input:
Title: SPATIAL VARIABILITY OF MICROZOOPLANKTON GRAZING ON PHYTOPLANKTON IN COASTAL SOUTHERN FLORIDA, USA
Review round: 2
Reviewer: 1

Output:
Basic reporting: The only further comments I have are about a small number of minor changes needed for the basic reporting:

line 51: fix typo here

line 134: is this meant to be "meta-analyses"?

line 201: change to "Data from the PCA were extracted"

line 227: change "Fig. 5" to "Fig. 3". Also please capitalise the word directly after this.

line 339: change to "dominant"

line 346: change "about" to "amount". Also, on line 348, add "is" between "it" and "surprising"

In the figures section, the captions in the page immediately before some of the figures is different to the caption added on the same page of each figure (I guess these should be the same). When this manuscript is further processed please make sure the correct/updated version of each caption is used.
Experimental design: no comment
Validity of the findings: no comment
Additional comments: No additional comments


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

Input:
Title: SPATIAL VARIABILITY OF MICROZOOPLANKTON GRAZING ON PHYTOPLANKTON IN COASTAL SOUTHERN FLORIDA, USA
Review round: 2
Reviewer: 2

Output:
Basic reporting: I'm glad to see that the authors have decided to include most of my suggestions in the manuscript. Not because I was the one making them but because I truly believe that they bring the manuscript a notch above. I am particularly happy to see the massive changes in the figures, in the calculation of photosynthetic and respiration data, and in the Supplementary Information. I also believe that the Discussion is now ready to put the author's novel results in a higher perspective and I like the flow of the text and the arguments presented. On the rare occasions where the authors decided not to include my comments, they have provided a sound rebuttal and I am happy with their responses and the references that support them.

In my opinion, the manuscript has been severely improved after the revision process and I am happy to have been a part of it. I honestly hope that the authors agree with me. Therefore, I recommend this manuscript to be accepted for publication in PeerJ as is.
Experimental design: No comment.
Validity of the findings: No comment.
Additional comments: No comment.


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: ASSOCIATION BETWEEN LARYNGOPHARYNGEAL REFLUX AND OBSTRUCTIVE SLEEP APNEA IN ADULTS
Review round: 1
Reviewer: 1

Output:
Basic reporting: - This paper examines the association between laryngopharyngeal reflux and obstructive sleep apnea syndrome (OSAS) in adults.
- The paper reports the following major findings:
1. Negative findings in most of the comparisons between the OSAA (mild-moderate and severe) and non-OSAS groups including:
 a). Mean Reflux Symptom Index (RSI),
 b). Mean Reflux Finding Score(RFS),
 c). The proportion of the laryngopharyngeal reflux (LPR),
 d). RSI and RFS correlation with Apnea-Hypopnea Index (AHI).
 e). The proportion of LPRs based on RFS between the Epworth Sleeping Scale (ESS) based groups.
2. Positive findings are:
 a). The proportion of LPR based on RSI varies significantly between Epworth Sleeping Scale (ESS) groups.

The paper clearly conveys the literature, methods, and major findings. Although there are multiple spelling and grammatical errors, for example, in the abstract, "index" is spelled "indeks." Proofreading can easily correct these minor mistakes.
Overall, the paper is self-contained with relevant results to support the claims.
Experimental design: Strengths of the paper
1. The paper includes clearly describes the introduction, methods, and major findings.
2. The paper performs sound statistical analysis and reports the details enough to replicate the study.
3. Conclusions are supported by the evidence presented in the paper.

Weaknesses

1. The paper includes the following statement in the conclusion section, “In conclusion, LPR is more related to decreased quality of life of 262 patients with OSAS than to cause apnea or hypopnea.” However, there is no evidence presented in the paper to support this claim. The paper infers this indirectly from ESS, which reflects average sleep propensity in daily life (ASP). To sustain this claim, the paper should include the Quality of Life (QoL) measure or exclude/change this statement as appropriate.
Validity of the findings: As the paper compares multiple tests and results, I would suggest correcting the p-values for the multiple comparisons to prevent false-positive results. Although it won't affect negative findings, it will rule out false-positive findings especially with the significant findings reported in the proportion of LPR based on RSI varies significantly between Epworth Sleeping Scale (ESS) groups.
Additional comments: Although negative findings are the primary findings of this paper, the paper involves sound methodology and gives enough details to be replicable. I recommend acceptance after 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: ASSOCIATION BETWEEN LARYNGOPHARYNGEAL REFLUX AND OBSTRUCTIVE SLEEP APNEA IN ADULTS
Review round: 1
Reviewer: 2

Output:
Basic reporting: No comment.
Experimental design: No comment.
Validity of the findings: No comment.
Additional comments: This paper deals with an important as well as an interesting topic, and is well-designed and well-written. The methodology and results are statistically robust. Some suggestions for improvement are stated below.

1)The Methodology within the Abstract should be presented in a more meaningful way. The study design is not clear as provided in its present form.

2) The statistical methodology should be stated in detail under a separate subsection within the Materials and Methods section.

3)The term “samples” should be replaced by “subjects” or “patients”.


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: ASSOCIATION BETWEEN LARYNGOPHARYNGEAL REFLUX AND OBSTRUCTIVE SLEEP APNEA IN ADULTS
Review round: 2
Reviewer: 1

Output:
Basic reporting: NA
Experimental design: NA
Validity of the findings: NA
Additional comments: My points raised during the previous review have been addressed by the authors. I have no further suggestions.


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: ASSOCIATION BETWEEN LARYNGOPHARYNGEAL REFLUX AND OBSTRUCTIVE SLEEP APNEA IN ADULTS
Review round: 2
Reviewer: 2

Output:
Basic reporting: No comment.
Experimental design: No comment.
Validity of the findings: No comment.
Additional comments: The revisions are satisfactory.


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: DO PATTERNS OF INSECT MORTALITY IN TEMPERATE AND TROPICAL ZONES HAVE BROADER IMPLICATIONS FOR INSECT ECOLOGY AND PEST MANAGEMENT?
Review round: 1
Reviewer: 1

Output:
Basic reporting: The authors provide and interesting and insightful study of insect mortality patterns at broad geographic scale.  However, aspects of this manuscript are difficult to understand, and there are some weaknesses in design that need to be addressed.
	To classify the studies in their data set, the authors use 39 degrees latitude as a dividing line between the tropics and temperate zone.  They state that this is derived from the Koppen-Geiger climate classification scheme.  However, the K-G scheme relies only on temperature, seasonality, and amount of precipitation.  It does not utilize latitude at all.  Why the authors chose 39 degrees latitude requires clarification.
	Designating a dividing line between the tropics and temperate zone by latitude alone will necessarily be a bit fraught and arbitrary.  However, 39 degrees is very far north to draw this line.  Traditionally, viewed only by latitude, the tropics begin at 23 degrees and the subtropics begin at 35 degrees.  In North America, where the majority of the studies in this data set occurred, any site (excepting coastal climates) north of 35 degrees experiences regular freezes in the winter.  Closer to 39 degrees, winters are severe.  In a study of insect mortality factors, the annual freeze line is the most biologically reasonable place to divide temperate from tropics as it should partially determine both abiotic mortality and the influence of the third trophic level on insect mortality.  I recommend either redefining your boundaries and re-analyzing the data accordingly or redefining your terminology.

Ln 34:  replace “for” with “from”
Ln 77: replace “for” with “than”
Ln 90:  Table S1 provides 51 not 56 studies from “tropical” locales.  In the Excel spreadsheet, 67 (not 56) of the 107 underlying studies lie south of 39 degrees latitude.
Ln 176 – 178:  Is this a test against all sources of natural enemy mortality combined?  Please provide the value for mean irreplaceable mortality of all natural-enemy factors combined.
Ln 185 – 188:  This statement is extremely confusing as written.  I cannot parse what you are trying to convey.  Please re-write and have an outside of your lab reader evaluate for clarity.
Ln 203 – 209:  This belongs in the Methods.
Ln 224 – 226:  I would be more convinced if you stress tested this result by seeing if it also applies to truly tropical sites: ie. those south of 23 degrees latitude.
Limitations and Insights:  This is a well written and insightful section.
Figure legends are insufficient and unclear and in the case of Figure 4, fragmentary.
Experimental design: Designating a dividing line between the tropics and temperate zone by latitude alone will necessarily be a bit fraught and arbitrary.  However, 39 degrees is very far north to draw this line.  Traditionally, viewed only by latitude, the tropics begin at 23 degrees and the subtropics begin at 35 degrees.  In North America, where the majority of the studies in this data set occurred, any site (excepting coastal climates) north of 35 degrees experiences regular freezes in the winter.  Closer to 39 degrees, winters are severe.  In a study of insect mortality factors, the annual freeze line is the most biologically reasonable place to divide temperate from tropics as it should partially determine both abiotic mortality and the influence of the third trophic level on insect mortality.  I recommend either redefining your boundaries and re-analyzing the data accordingly or redefining your terminology.
Validity of the findings: See above
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: DO PATTERNS OF INSECT MORTALITY IN TEMPERATE AND TROPICAL ZONES HAVE BROADER IMPLICATIONS FOR INSECT ECOLOGY AND PEST MANAGEMENT?
Review round: 1
Reviewer: 2

Output:
Basic reporting: This manuscript meets all requirements for basic reporting. It is is clearly written and it follows PeerJ guidelines.
Experimental design: The methods used for this review are original, first developed by the same authors group and to me they seem to be adequate and well described.
My only recommendation will be to briefly state which were the criteria to select the papers used in the review. I understand that they are in the references, but it would be easier for the reader if they were listed  in the text.
Validity of the findings: The results are very interesting and address issues that have not been tested in ecological theory. The approach seems original to me and the literature support for the analysis is outstanding.
Additional comments: This manuscript represents an impressive review of the literature on life table studies (107 publications, 268 life tables). Causes of death were analyzed and irreplaceable mortality by cause was estimated using multiple decrement life tables. Comparisons are made between temperate and tropical zones, as well as between native and non-native species. The research work is a logical continuation of the work by Peterson et al (2009), where applying the same methods expands the database to make this type of comparisons. In my opinion this is an excellent paper. The Multiple Decrement Life Table analysis provided as supplementary material is extremely valuable.
The results challenge general ecological expectations, such as the greater importance of abiotic factors in temperate climates, or the greater susceptibility of native insects, compared to non-native ones, to natural enemies. However, these general expectations have not always been empirically validated. For me this highlights the importance of this study and for that reason I recommend accepting it for publication.

Specific comments.
L 82. “… so we can compare” instead of “… “so we are able to compare”.
L 104-105. Could you state which were those basic criteria? I understand that they are in the references, but it would be easier for the reader if they were listed here. For example, I assume you only consider open field life-table studies and do not consider laboratory experimental studies.
L 177-178. Include the mean percent (±SE) of irreplaceable mortality for the "all types of natural-enemy mortality" factor. According to figure 2 is aprox. 21%
L 187-188. Delete “which were all natural enemies, all parasitoids, pathogens, and all non-natural enemy factors”. There is no need to repeat.
L 210-220. I am not sure about the implications of this cluster analysis. Does it really have a biological meaning? Honduras is a tropical country and is in cluster 2.  Is hard to explain why in Canada and Kenya, pathogens have a more relevant role than in other locations. Perhaps you might reconsider to delete this non-hierarchical multivariate k-means clusters analysis.
L 224 – 226. Could this be explained because this category includes "other causes" or not identified causes? For example, the cause of death might be a pathogen, but without symptoms that allow its identification. Or the cause of death might be physiological, intoxication or other cause that in insect life -table studies are not considered.
L 316-316. This recommendation should be taken with caution, because in tropical zones predators generally are already present. The introduction of predators can cause great disturbances. Perhaps conservation measures.
L 317-319. The evidence on the effect of parasitoids producing high levels of parasitism (= mortality) is extensive, although not in life table studies. Do you think this could be an explanation for the low mortality caused by parasitoids?
Legend of Figure 4 was cut or is incomplete


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: DO PATTERNS OF INSECT MORTALITY IN TEMPERATE AND TROPICAL ZONES HAVE BROADER IMPLICATIONS FOR INSECT ECOLOGY AND PEST MANAGEMENT?
Review round: 2
Reviewer: 1

Output:
Basic reporting: Ok.
Experimental design: The authors have addressed my previous criticisms.   Although I still think more consideration could be given to dissecting the temperate – tropical “dichotomy”, and I think the clustering based on political boundaries is problematic (see below).  On balance I think this is a worthy study.

The K-means clustering of countries based on the studies conducted therein is a bit troubled.  The authors include all countries in the study, however many of these countries are only represented by 1 to 3 lifetable studies.  It is stretch to state that these few studies present a framework from which to generalize at the country level.   In fact, most of the countries in these clusters are underpinned by 5 or fewer lifetables.  Probably these countries should be dropped from the analysis.  (And the 2 studies from Hawaii, should probably not be lumped with studies from the “United States”.)  Alternatively, a more logical approach in my view, would be to group studies by biogeography rather than political boundaries.

93 – 96:  In fact, you did include a sub-tropical group, you just did not split it out.  Studies from the subtropics constitute a little over 1/3 of your tropical group.  This raises the question of how reflective your results are of tropical systems.  It was for this reason that I previously recommended stress testing your results by conducting an analysis restricted to those studies that took place in unquestionably tropical versus clearly temperate environments.  If the patterns hold, even if you lack sufficient power for significance, the reader can have more confidence in your findings.  This approach would be particularly important for results that are surprising.  For example, the lack of difference in irreplaceable mortality for parasitoids or the overall balance of abiotic and biotic mortality.  Do these values shift in the direction predicted by existing theory when a more tropical set of life tables is examined?
Validity of the findings: The authors have addressed my previous criticisms.   Although I still think more consideration could be given to dissecting the temperate – tropical “dichotomy”, and I think the clustering based on political boundaries is problematic (see below).  On balance I think this is a worthy study.

The K-means clustering of countries based on the studies conducted therein is a bit troubled.  The authors include all countries in the study, however many of these countries are only represented by 1 to 3 lifetable studies.  It is stretch to state that these few studies present a framework from which to generalize at the country level.   In fact, most of the countries in these clusters are underpinned by 5 or fewer lifetables.  Probably these countries should be dropped from the analysis.  (And the 2 studies from Hawaii, should probably not be lumped with studies from the “United States”.)  Alternatively, a more logical approach in my view, would be to group studies by biogeography rather than political boundaries.

93 – 96:  In fact, you did include a sub-tropical group, you just did not split it out.  Studies from the subtropics constitute a little over 1/3 of your tropical group.  This raises the question of how reflective your results are of tropical systems.  It was for this reason that I previously recommended stress testing your results by conducting an analysis restricted to those studies that took place in unquestionably tropical versus clearly temperate environments.  If the patterns hold, even if you lack sufficient power for significance, the reader can have more confidence in your findings.  This approach would be particularly important for results that are surprising.  For example, the lack of difference in irreplaceable mortality for parasitoids or the overall balance of abiotic and biotic mortality.  Do these values shift in the direction predicted by existing theory when a more tropical set of life tables is examined?
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: EXCITO-REPELLENCY OF MYRISTICA FRAGRANS HOUTT. AND CURCUMA LONGA L. EXTRACTS FROM SOUTHERN THAILAND AGAINST AEDES AEGYPTI (L.)
Review round: 1
Reviewer: 1

Output:
Basic reporting: Dear Authors
Overall study is scientifically sound but has shortcoming to incorporate before consideration.
About title
I think that this is unsuitable to state here because study id repelling the A. egypti....... Please change this in the title replacing vector with mosquito name.......
Introduction
Please make the objective more clear by adding the data of repellent background in the last paragraph.
please add the more data of the biochemical potential of plant extracts..... which will be actual background of the study...
Line 49-51: Please make two sentences and rewrite it.
Line 118-119: Move in the materials and methods.
Materials and Methods
Line 123: change was with were
In the discussion the comparison of previous studies is not bad but it can be accepted somehow...... Authors almost have more than half discussion of this nature which i do not think a good in publication...... Please make the discussion good according to the trends of your results....... Discuss your plants extract potential/efficacy rather than discussing other plants......
Line 279-280: Above in introduction you have mentioned that this extracts has never been tested before?????
In the conclusion, Please conclude some results.
Experimental design: Overall Methods of study is explained in considerable but some queries are still to answer
Why you did not use the mean comparison tests (Tuckeys, Dunnetts etc.), while you have different treatments groups.....
I do not think that without theses authors can compare which is better one.......
Where is replication scheme???
if pseudo replication then how authors manage the experimentation????
Roberts et al. 1997 methods: Please explain here and cite at the end of text........
Validity of the findings: Overall the results are satisfactory....
On more analysis must be in the results section......
mean comparison test using tuckeys test.....
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: EXCITO-REPELLENCY OF MYRISTICA FRAGRANS HOUTT. AND CURCUMA LONGA L. EXTRACTS FROM SOUTHERN THAILAND AGAINST AEDES AEGYPTI (L.)
Review round: 1
Reviewer: 2

Output:
Basic reporting: Overall study of this paper is good but I am concerned that the paper should be improved to meet the standard of PeerJ for publication. The manuscript in its current form is difficult to considered for publication, as it need to resolve suggested changes.
Title is not adequate for the content and given information of this paper. It must be precise and clear according to the data mentioned in methodology.
Abstract is written well but need addition of future directions.
Introduction data is not sufficient to discuss the background of the designed study. Please add more relevant text to improve the reading context of the title studied. Remove old references. The hypothesis should be clearly stated in the introduction, which can never be modulated with a strong story background of the study. It is suggested to improve it.
L 80-82: Need to rephrase this sentence and citations must be at the end of this sentence.
L 83-86: Citation at the end of sentence.
L 97:  DEET, explain this word
L 118 and 119: No need to mention figure here.
The authors need to precise the data in discussion and remove old references.
Experimental design: The experimental design of this study is well explained. Need few changes i.e.,
L 183: 24-h “24 h”
L 185: 3-min “3 min”
L 189: 30-min “30 min”
Validity of the findings: The results are good for the presentation of this study.
Additional comments: I would like to commend the authors for their efforts, but there is need to make the paper interesting for an international audience such as the PeerJ readers.


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: EXCITO-REPELLENCY OF MYRISTICA FRAGRANS HOUTT. AND CURCUMA LONGA L. EXTRACTS FROM SOUTHERN THAILAND AGAINST AEDES AEGYPTI (L.)
Review round: 1
Reviewer: 3

Output:
Basic reporting: It is an interesting study having a sufficient research data and well explained results in an effective way. More interestingly, it is the first study in this field that illustrate the effect of two plant materials on the zika virus. Here are few minor changes in the research article Please rectify all with keen concern to improve the paper.
It seems there is need to rectify the font size of all headings, also make it sure that all headings and sub-headings have same font. “Justify” all paragraphs in the manuscript. Furthermore, many grammatical errors have been observed in the manuscript, please read it again carefully and remove all obstacles. Sentence making also need to improve to clarify the central idea of the research, as at some points in the paper it creates ambiguity in the actual meaning of sentence due to poor sentence making. Objectives of this research are unclear, Hence, there is need to more explanation in comprehensive way of research objectives in the end of “Introduction”. Figures are unnecessary in the introduction, please add research relevant plants figures in the materials and methods if seems necessary. Few suggestions are following:
1-	line 47 of introduction replace mosquitos to mosquito
2-	line 48 also give other examples of “several tropical disease pathogens”
3-	Line 51, Remove “Is Concerning”
4-	Line 52 Remove “At least”
5-	Line 53 Replace is >Was
6-	Line 56 Replace till date>Until now
7-	Line 57 Replace To be >is
8-	Line 65 Remove “and” before readily available
9-	Line 67 Remove “Since”
10-	Line 69 replace Can be >Could be
11-	Line 73, 74. There is no need for any figure in introduction. You might include it in the Materials and Methods if it would be the part of your Research work.
12-	Line 76, remove “Symptoms”
13-	Line 90, No need for any figure in introduction
14-	Line 98 Replace Rhizome essential oils>its essential oil
15-	Line 118 No need of figures in introduction
16-	Line 118-119 seems unfit in introduction, must be the part of materials and methods.
Experimental design: This part is very well explained with adequate detail of each step of the experiment. Meanwhile, few minor points need to edit to improve the significance of paper. Please add references of all protocols used in this research. “Data analysis” part of this section should be at the end of materials and methods. It would be more beneficial for better understanding if number of plants also mentioned in this section. Figures of plants and extracts used in research material should add in this part. Few more points are following:
1-	Line 142 remove “and” before mace
2-	Line 166 remove “or”
3-	Line 179 replace are >were
4-	Line 180 replace is >was
5-	Line 187 replace was>were
6-	Line 191 replace was>were
7-	Line 196 to 206 must be the last lines of Materials and Methods. For this purpose shift the complete heading data of “Data analysis” after the “Gas chromatography-mass spectrometry (GC-MS) analysis”.
Validity of the findings: Research results are well explained, and novelty of the research is well describe in this section in a brief way to meet all requirements of research objectives. Albeit on, this section needs more concentration on correct sentence making and improvement in grammar as well. Please use less quantitative values to support  the results in discussion, it creates cliché. Some points need more justification to support the results
1-	Lines 250-257 required more justification/references to support the results
2-	Line 334 needed more references to support the results
Additional comments: I have keenly review the manuscript and try my best to fulfil the general requirements of the journal. It is an interesting study having all the essential requirements of a good paper especially, the authors have tried to manipulate the new research area in best possible way.


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: EXCITO-REPELLENCY OF MYRISTICA FRAGRANS HOUTT. AND CURCUMA LONGA L. EXTRACTS FROM SOUTHERN THAILAND AGAINST AEDES AEGYPTI (L.)
Review round: 2
Reviewer: 1

Output:
Basic reporting: I suggest the the title below because it is still not looks very impressive
"Excito-repellency of Myristica fragrans Houtt. and Curcuma longa L. extracts against Aedes aegypti (L.) in Southern Thailand"
You may choose any other on your choice.
Experimental design: NA
Validity of the findings: NA
Additional comments: NA


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: EXCITO-REPELLENCY OF MYRISTICA FRAGRANS HOUTT. AND CURCUMA LONGA L. EXTRACTS FROM SOUTHERN THAILAND AGAINST AEDES AEGYPTI (L.)
Review round: 2
Reviewer: 2

Output:
Basic reporting: The authors resolved all the suggested changes that I have mentioned previously. In present form the paper is according to the standard of PeerJ for publication. I appreciate to the authors that they did a lot of hard work for this study.
Experimental design: N/A
Validity of the findings: N/A
Additional comments: N/A


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

Input:
Title: EXCITO-REPELLENCY OF MYRISTICA FRAGRANS HOUTT. AND CURCUMA LONGA L. EXTRACTS FROM SOUTHERN THAILAND AGAINST AEDES AEGYPTI (L.)
Review round: 2
Reviewer: 3

Output:
Basic reporting: This study is scientifically clear and advanced knowledge have been given in the article.  The background knowledge is also sufficient and no ambiguity found in the data. Results are according to the hypothesis and fulfil the requirements of the study. Author has provided enough data to publish this manuscript in the PeerJ.
just Minor change is recommended:

L96: Replace “Curcuma longa”> C. longa
L122-123: Remove “To the best of our knowledge, the ER properties of M. fragrans 123 and C. longa extracts have never been accurately studied” as it is repeated in next paragraph with more detail.
Experimental design: Research question is well defined and relevant to the study. Methodology is also clearly explained with sufficient information about the research.
Validity of the findings: All provided data is statistically sound and robust. conclusions is also well stated and linked to the primary research objective.
Additional comments: N/O


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: RFPDR: A RANDOM FOREST APPROACH FOR PLANT DISEASE RESISTANCE PROTEIN PREDICTION
Review round: 1
Reviewer: 1

Output:
Basic reporting: In general, the work is solid but routine judged from this type of work in the field of bioinformatics. The manuscript is fairly clear.
Experimental design: 1)	Some baseline methods (such as SVM, Logistic Regression) should be tested based on the dataset and feature vectors used in this work. By doing so, the readers will have global understanding of different ML algorithms in this topic.
2)	It is also necessary to compare the proposed method against PSI-BLAST (i.e., Sequences in testing set should be PSI-BLASTed against training set. If the label of top hit is the same as the query sequence, a correct prediction is obtained; otherwise, it is a wrong prediction). By doing so, the readers will know the advantage of the proposed alignment free method in comparison to sequence alignment-based method.
3)	The choice of P:N ratio seems to be an open issue in this type of bioinformatics task. In real application, which ratio should be used for training models?
Validity of the findings: No comment.
Additional comments: 1) Line 98, the threshold used for sequence redundancy removal should be provided.
2) Line 153-158, the implementations of RRGPredictor and DRAGO2 should be provided. Were the benchmarking experiments done through the corresponding web servers or software for local use?
3) Table 4, what are the thresholds for these three methods? It seems that ROC curves or PRC curves are more informative to compare these three methods comprehensively.
4) Table 4, the computational time means the time for all predictions or one prediction? Moreover, the computer configuration should be mentioned when talking about the computational time.


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: RFPDR: A RANDOM FOREST APPROACH FOR PLANT DISEASE RESISTANCE PROTEIN PREDICTION
Review round: 1
Reviewer: 2

Output:
Basic reporting: More details in general comments section
Experimental design: More details in general comments section
Validity of the findings: More details in general comments section
Additional comments: In this manuscript, authors used a random forest approach to predict disease resistance (DR) genes in plants. This approach is novel, since previous studies were using SVM and using a much smaller training set. Overall, the analysis done is solid and the manuscript is well written. However, I am concerned whether ML is really needed, and whether existing approaches are good enough. There also needs to be a better discussion of the results on why some features are important. My detailed comments are below:

Major comments:
Introduction – authors should give more explanation for the diversity of DR genes. For an uninformed reader, this will not be clear. Also, is DR the same as Pathogen Recognition Genes (PRGs) used in Osuna-Cruz, 2018 and in PRGdb? Why not use the same abbreviation? If it is called DR, there are multiple mechanisms of disease resistance – are the authors trying to predict genes involved in all of these mechanisms? Clarify only.

Are all proteins in the non-DR set really non-DR? Authors only removed proteins with PFAM domains previously mentioned in Osuna-Cruz et al, 2017. Does that mean DR proteins can simply be found by searching for those PFAM domains? The problem of defining a true negative set for ML is a real serious problem in biological ML.

L164: Why was the Mann-Whitney-Wilcoxon test used only for tripeptides and not for other features? Appropriate feature selection must be performed even for other features, since there are too many features for the number of training examples, leading to overfitting. Authors should carefully consider if all these features are really needed for DR protein prediction.

Supplementary Table S1: Authors used an fdr p-value threshold of 0.05 to select tripeptide motifs. I think since the comparison is between 500 positive and 68,000 negative examples, having 5 instances in positive and 0 instances in negative is also going to show up as very significant. Authors should try to filter using more stringent threshold or some threshold based on frequency. Most features have minimal effect on model performance anyways.

Fig. 3/Table 2 – the high performance using all strategies indicates some existing trivial structure to the positive and negative training sets. I am not sure what it is, but my hunch is the filtering for PFAM domains that the authors did for the negative set. The positive set must then contain PFAM domains with unique structures not present in the negative set. Thus I feel the problem could be simply solved by knowing the domain architecture.

Fig. 4: To complement this analysis, authors should also look at the top 10-20 features and determine the performance metrics if only those features are used, and present it as Supplementary. As presented here, the analysis seems to be a black box, and why the top features are important is not clear. Authors provide some discussion on lines 280-288, but it is only for some tripeptide motifs. What about CTD, transition and composition, which are the most important feature sets?

On line 268, authors mention Moreau-Broto autocorrelation and conjoint triad as the most frequent categories. What are those? Why not some other autocorrelation? Looking at Table S2 further confused me on what the individual features really correspond to in terms of protein structure.

Table 4 – it is not clear if the performance of this method is really better than previous methods. Why not just use DRAGO2 or RRGPredictor? The authors should also compare their performance to that using simply PFAM/Interpro domain information. From the RRGPredictor paper, it appears that there are 10 standard domains in DR proteins. Would simply using Interproscan to find these domains be equally as good as this?

Are there new types of DR genes identified using this approach that RRGpredictor or PRGdb are not able to identify?

In the Discussion, authors mention that Interproscan takes time, but most genome databases already provide domain information. Softwares like hmmscan also run within a matter of minutes on entire plant proteomes using multi-processor architectures. Furthermore, I believe using RD-RFPDR would require calculating all features for all proteins for each genome using protr, which would also be a time-consuming step, comparable to hmmscan/IPRscan. So I am not sure if this is a valid argument.

Minor comments:
Abstract: While the rest of the manuscript is written properly, there are some grammatical mistakes in the abstract. E.g. “are among the”, “in order to generate also a”

Lines 27-224 where authors describe the importance of different features should be moved to the introduction. Why not also use domain-related features?

Fig. 3 is only briefly mentioned in the Results, and although these results are in Discussion, the figure is not cited. Please add the citation in Discussion.


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

Input:
Title: RFPDR: A RANDOM FOREST APPROACH FOR PLANT DISEASE RESISTANCE PROTEIN PREDICTION
Review round: 2
Reviewer: 1

Output:
Basic reporting: No  comment.
Experimental design: No comment.
Validity of the findings: No comment.
Additional comments: Thanks for the authors' efforts to carefully address my comments. I would like to suggest  the acceptance.


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

Input:
Title: RFPDR: A RANDOM FOREST APPROACH FOR PLANT DISEASE RESISTANCE PROTEIN PREDICTION
Review round: 2
Reviewer: 2

Output:
Basic reporting: The authors have addressed all my concerns, and the manuscript looks good. Despite Table S3, I would still add some information about the different features, especially the unusual autocorrelation features, in the Intro/Results/Discussion. I would also recommend splitting the Discussion into more paragraphs. These are optional recommendations.
Experimental design: No comment
Validity of the findings: No comment
Additional comments: No 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: PREVALENCE AND INCIDENCE OF SARCOPENIA AND LOW PHYSICAL ACTIVITY AMONG COMMUNITY-DWELLING OLDER THAI PEOPLE: A PRELIMINARY PROSPECTIVE COHORT STUDY 2-YEAR FOLLOW-UP
Review round: 1
Reviewer: 1

Output:
Basic reporting: The research question is well warranted and relevant. It is also well-defined and meaningful to the older adult population. Well done on examining an area that has been underdeveloped and bringing to fruition an area that will make a significant impact.  Major revisions are required as I believe there are sections that need to be more clear and concise, given the importance and significance you state with this research topic. This message of clarity does not come across yet, especially being that this is the first study to investigate sarcopenia in Thia older adults residing in the community.

The areas of interest that need editing are the following:

In terms of your title, recommend removing the COVID-19 pandemic as you do not mention COVID-19 anywhere else in your manuscript.

The reference list needs to be screened through as currently does not follow a consistent PeerJ style.

Table formatting needs to be cleaned up (provided comments in the attachment).
Experimental design: Materials and methods
How many participants were invited to participate and screened in 2019, this was not stated in the manuscript
In terms of the walk test, what did participants do to complete this? using what equipment and instructions were provided to the participants - as you include relevant information for handgrip strength and BIA but this is missing for the walk test.

In the results you mention severe sarcopenia please provide in the introduction what this terminology means and the significance of this term.
Validity of the findings: The discussion needs some work on the delivery of the results and the importance of the study being brought up earlier (see attached file). the relevance of stating that this is the first study to explore the incidence of sarcopenia among Thai older adults needs to be driven and consistent throughout the manuscript.
Additional comments: Please see the attached file that has been scanned which provides more comments around spelling, grammar, tense and edits to your 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: PREVALENCE AND INCIDENCE OF SARCOPENIA AND LOW PHYSICAL ACTIVITY AMONG COMMUNITY-DWELLING OLDER THAI PEOPLE: A PRELIMINARY PROSPECTIVE COHORT STUDY 2-YEAR FOLLOW-UP
Review round: 1
Reviewer: 2

Output:
Basic reporting: This manuscript is written well, however, this need to be revised a few points.
1) Please describe the hypothesis of this study more specifically.
2) Figures are hard to understand. Also, it is not clear. Please correct.
Experimental design: no comment'
Validity of the findings: no comment'
Additional comments: no comment'


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: PREVALENCE AND INCIDENCE OF SARCOPENIA AND LOW PHYSICAL ACTIVITY AMONG COMMUNITY-DWELLING OLDER THAI PEOPLE: A PRELIMINARY PROSPECTIVE COHORT STUDY 2-YEAR FOLLOW-UP
Review round: 1
Reviewer: 3

Output:
Basic reporting: 1. The lines in Figure 1 should be revised to be standard.
2. Line 160-162 cannot be supported by Table 3, please explain or revise.
3. The language should be revised throughout the manuscript with help of native English speakers.
Experimental design: 1. As shown in line 111-114, Sarcopenia diagnosis and classification should be checked and corrected. Based on AWGS 2019, severe sarcopenia is diagnosed as low handgrip strength + slow gait speed and low SMI. Your definition of possible sarcopenia is also incorrect.
2. As the physical activity is mostly associated with novel findings of this study, more details regarding the assessed results by Global Physical Activity Questionnaire should be reported, such as exercise program, intensity, and frequency of included participants.
3. The authors only reported changed prevalence rates of sarcopenia in two time points, however, the significance of these difference should be assessed with appropriate statistic methods, such as McNemar tests. Besides, the changes in handgrip strength, gait speed and SMI across two time points should also be assessed using paired sample T tests or Wilcoxon tests.
Validity of the findings: no comment
Additional comments: This study was well designed but with a small sample. Although several important findings were reported, such as the increased prevalence of sarcopenia and the roles of physical activity in reversing sarcopenia, these outcomes are in line with expectations. The manuscript may be accepted for publication if the listed problems be elegantly solved.


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: PREVALENCE AND INCIDENCE OF SARCOPENIA AND LOW PHYSICAL ACTIVITY AMONG COMMUNITY-DWELLING OLDER THAI PEOPLE: A PRELIMINARY PROSPECTIVE COHORT STUDY 2-YEAR FOLLOW-UP
Review round: 2
Reviewer: 1

Output:
Basic reporting: No further comments. Well done!
Experimental design: No further comments.
Validity of the findings: No further comments.
Additional comments: No further comments.


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

Input:
Title: PREVALENCE AND INCIDENCE OF SARCOPENIA AND LOW PHYSICAL ACTIVITY AMONG COMMUNITY-DWELLING OLDER THAI PEOPLE: A PRELIMINARY PROSPECTIVE COHORT STUDY 2-YEAR FOLLOW-UP
Review round: 2
Reviewer: 2

Output:
Basic reporting: Well done.
Experimental design: Well done.
Validity of the findings: Well done.
Additional comments: The authors have well revised the manuscript based on 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: AN EARLIEST TRIASSIC AGE FOR TASMANIOLIMULUS AND COMMENTS ON SYNCHROTRON TOMOGRAPHY OF GONDWANAN HORSESHOE CRABS
Review round: 1
Reviewer: 1

Output:
Basic reporting: In general, the idea of scanning rare fossil material in the synchrotron is good as it might reveal internal structures otherwise not accessible. However, the authors produced 3D surface models (which is fine), which they then compare with “normal” photos of different quality, especially concerning the light settings (see below). They then emphasise that with their method, structures become visible which are not accessible with normal photographic techniques, which I doubt for most cases. In my own experience, especially also with fossil xiphosurans, it is possible to access details of the surface structures with normal photographic equipment, which even allows stereo imaging (see also references mentioned below). Therefore, the authors should not compare their 3D models with normal photographs, but with stereo images of the same specimens.

Additionally, the manuscript has a second focus on the age of one of the species. However, there is no clear explanation in the methods part how the authors came to these conclusions, but a very lengthy geological context section, in which it does not become clear, which results are their own ones and which ones were already known previously (see also details below).

I think that this manuscript could be published after major revision.
Experimental design: Geological context (lines 128-213): This entire section, which is about 3.5 pages long, is not mentioned in the abstract at all. Also, I am not sure what the authors want to tell with this section. Apparently, these data are already mostly known as many references are cited here. Also, this section does not contribute much to the topic of the paper. Therefore, I suggest to delete most of it ans to only point out the aspects important for the context of this study.
The part on the age of Tasmaniolimulus would probably be interesting to keep, but in the Results section and more focussed on which aspects are new, so what the authors have actually done and found out. There is also no explanation in the material and methods section what the authors did to find our about the age of the specimens.

3D reconstructions: I think it would be necessary to compare the virtual 3D reconstructions, which the authors show in their figures, with simple stereo images taken from two different angles. In my experience, these simple images provide very good 3D information (e.g. as red-cyan stereo anaglyphs) and can be produced with simple camera equipment or even a flatbed scanner. Additionally, such simple stereo images do not go through an interpretation process, in contrast to the production of the 3D reconstructions, which the authors did.

Figures: It is stated that “plain light” was used. However, it is clear that in Fig. 1A and 2A (for example, but also in some other images) the lighting was set differently. In Fig. 2A, the light was to a certain extent directed, which is why shadows are visible. Therefore, the appendages are well visible in Fig. 2A (maybe even more clearly than in Fig. 2B). The authors need to rephrase the captions to make clear, which lighting setting was used in the different images, especially as they are pointing out the differences in the methods. They should also provide a statement about the virtual lighting for the renders of the models. If, for example, a ray-tracing algorithms was used, the influence of the lamps can be quite drastic.

General comment: The term horseshoe crabs, though common, is rather unfortunate as these animals are not at all closely related to crabs (which the authors are clearly aware of). Better change to Xiphosurida or whichever other name is preferred.
Validity of the findings: see other comments
Additional comments: 24: Though synchrotron CT (and also other CT types) are often termed non-destructive, this is not really the case. The method causes some destruction, though not much, depending on the type of fossils or on the mounting method etc. Please rephrase here and in other places where necessary.

35: Use Euchelicerata instead of Euchelicerate

47: Use Metazoa instead of Animalia, which is rather outdated

63-67: I would simply delete these brackets [ ] as they decrease the readability.

69-70: You wrote “most 3D data collected from fossil xiphosurids have been surface scans”, but cite only one paper, please include some more references here. In general, normal stereo imaging is not uncommon in palaeontological papers and would fit here. Also, I do not want to do citation pushing, but a suitable paper would be this one (especially check fig. 3e):
Haug, C. & Rötzer, M. A. I. N. 2018. The ontogeny of the 300 million year old xiphosuran Euproops danae (Euchelicerata) and implications for resolving the Euproops species complex. Development Genes and Evolution 228, 63–74.

86: Change to “species of Xiphosurida”.

91: Change “taxa” to “species”

92: I recommend to avoid Linnean ranks such as families (rank height is very subjective). Here, it could simply be substituted by ingroups.

98: cm is no SI unit, use mm instead.

129: Use Australian instead of Australia

184: I think it should read “sufficiently”?

226: Use species instead of taxon (also check in other places).

214 ff.: The whole results section is very short. Maybe some more description would make it easier for the reader to follow.

231: As said above, it is not clear how the authors found out about the new age (that means, which methods they used here, that should be in the methods section).

231: Why does the revised age have “important implications for austrolimulid evolution”? The age of fossils does not directly affect the phylogenetic relationships, and oldest fossils do not necessarily have the most ancestral morphology. Please explain what you mean here, the following sentences are not really sufficient here.

252-254: Again, I do not aim at pushing own papers, but as the authors mention that “there are limited examples”, these two show very good examples for ventral preservation (which are also shown as stereo images):
1) Haug, C. & Rötzer, M. A. I. N. 2018. The ontogeny of the 300 million year old xiphosuran Euproops danae (Euchelicerata) and implications for resolving the Euproops species complex. Development Genes and Evolution 228, 63–74.
2) Haug, C., Van Roy, P., Leipner, A., Funch, P., Rudkin, D. M., Schöllmann, L. & Haug, J. T. 2012. A holomorph approach to xiphosuran evolution—a case study on the ontogeny of Euproops. Development Genes and Evolution 222, 253–268.

275: carapace is a tricky term, better use prosomal shield

298-301: But CFD has also been used, e.g., for ammonites: https://palaeo-electronica.org/content/2020/3003-cfd-of-ammonoid-shells
What about CFD studies on large marine crustaceans, are there any?

Caption of Fig. 4: Reference to C is missing, instead F is referred though not in figure, needs to be corrected.


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: AN EARLIEST TRIASSIC AGE FOR TASMANIOLIMULUS AND COMMENTS ON SYNCHROTRON TOMOGRAPHY OF GONDWANAN HORSESHOE CRABS
Review round: 1
Reviewer: 2

Output:
Basic reporting: The paper is mostly well-written, with sufficient background and references to the relevant literature provided. There are a number of typos in the text, and in the attached annotated manuscript, I have made corrections and suggested some rephrasing of sentences in sticky notes. The figures are sufficient, of good quality, and backed up by further supplementary images uploaded online.

However, in some cases, from the text, it is not entirely clear what the authors are trying to say. For a detailed discussion of these issues, please see the "Additional comments", and the annotated manuscript.
Experimental design: So far, only one single fossil xiphosurid had undergone synchrotron CT scanning. So, the four specimens scanned by the authors definitely represent a welcome addition. The method and its research potential are sufficiently explained, and this part definitely meets the requirements set by PeerJ.
Validity of the findings: Some of the discussion and conclusions are a bit muddled, in part due to rather imprecise phrasing, which makes it unclear what exactly the authors are trying to say (e.g. "cuticularised", "ventral preservation"). Also, while the text states that the CT imaging revealed moveable spines in Victalimulus mcqueeni, only the fixed spines, and intervening notches are visible in the images provided. For a detailed discussion of these issues, please see the "Additional comments", and the annotated manuscript.
Additional comments: Review of “An earliest Triassic age for Tasmaniolimulus and comments on synchrotron tomography of Gondwanan horseshoe crabs” (#67956)

	Horseshoe crabs are an iconic clade of euarthropods with a long evolutionary history, and the only extant fully aquatic chelicerates. While currently only four species are in existence, their taxonomic diversity and morphological disparity used to be much greater in the Paleozoic and Mesozoic. Therefore, any paper dealing with fossil xiphosurids is of considerable interest to palaeobiologists and invertebrate and evolutionary biologists.

	The current manuscript revises the age of Tasmaniolimulus, previously believed to be Permian, to the Triassic, which has implications for our understanding of the timing of the radiation of the family Austrolimulidae. In addition, it discusses the results of synchrotron CT scanning of four specimens of fossil Australian horseshoe crabs. The extremely high energies, brilliance and almost monochromatic character of synchrotron light allow a penetration and resolution unequalled by classical X-ray tomography. Therefore, synchrotron tomography is quickly becoming established as an extremely valuable technique in the study of fossil specimens. However, so far, synchrotron tomography had only been applied to one fossil horseshoe crab specimen. The four specimens scanned for this paper therefore represent a welcome addition, and provide some additional morphological data over what was known previously from more classical studies of the material using imaging in visible wavelengths. Consequently, this paper is of interest, and deserves to be published. There are, however, a number of issues that need to be addressed first.

	On p. 14, line 221, the authors state that “... a limited portion of the spine was cuticularised.” What is this supposed to mean? What is “cuticularised”? Xiphosurids are euarthropods. Euarthropods are Ecdysozoa. Ecdysozoans, as one of their primary defining characters, have an external cuticle. In euarthropods, most of this cuticle (except for arthrodial membrane) is at least to some degree sclerotised to form an exoskeleton. So, a euarthropod, by definition, is externally encased in a sclerotised cuticle. I suppose the authors may be trying to say to say that the degree of sclerotisation varied along the spine, or that the whole spine was less sclerotised compared to the rest of the prosomal shield? This sentence does need rephrasing to make sense – talking about a portion of the external surface of an ecdysozoan being “cuticularised” is akin to talking about a portion of a mammalian spine being “notochordised”.

	On p. 15, lines 227-228, it is stated that “The reconstruction of Victalimulus mcqueeni reveals evidence for the thoracetronic doublure, moveable spines and notches, and appendage impressions.”
-	First, after reviewing both Fig. 4 and the supplemental image, I fail to see the moveable spines – I can only make out the fixed spines of the thoracetron, and the intervening notches that accommodated the moveable spines, not the moveable spines themselves. Indeed, the caption to Fig. 4 itself states: “... X-ray tomographic slice showing fixed spines and moveable spine notches (white arrows) and thoracetronic doublure”; so the caption itself does not mention the presence of the actual moveable spines either. If the moveable spines are indeed visible anywhere, they should be indicated.
-	Second, the statement is somewhat misleading, considering that the original description by Riek & Gill (1971) already commented on the thoracetronic doublure, the presence of fixed spines and notches, and the impressions of the prosomal walking limbs. Therefore, these do not exactly represent new findings.

On p. 16, the authors talk about “dorsal preservation”, and “ventral preservation”. It is not entirely clear what they mean by this:
-	Does it mean that only the dorsal (or ventral) exoskeleton is preserved?
-	Or, do the authors intend to say that the fossils are exposed from the dorsal (or ventral) side?
-	Or, do they intend to convey that the original orientation in which the specimens were preserved was with the dorsal (or ventral) side up?
These are not trivial differences in meaning. The authors comment that “dorsal preservation” is more common than “ventral preservation” in horseshoe crabs. If they are referring to the surface of the fossil that is exposed, this is not surprising: the dorsal surface represents a relatively smooth, convex surface, whereas the concave ventral surface is much less regular. As such, the dorsal surface will represent a plane of weakness along which the matrix containing the fossil will preferentially fracture. It therefore makes sense for the majority of xiphosurid specimens to be exposed from the dorsal side – as is observed for most other dorso-ventrally flattened euarthropods. However, the fact that these specimens are exposed on the dorsal side does not necessarily mean that they have been preserved dorsal side up: it is common for specimens which have been embedded dorsal side down to still crack out along the dorsal surface, because of the aforementioned reasons. Hence, if it is the intention of the authors to make the case that most horseshoe crab fossils were preserved with the dorsal side up, they need to provide additional arguments for the original orientation of the specimen – either from sedimentological characteristics of the surrounding matrix that allow polarity to be deduced, or from data documenting the original orientation of the fossil when collected.

	On the same page, the authors also comment that “... a ventral orientation has a lower preservational protentional”. Again, if they are just referring to the side of the fossil that is exposed, it makes sense, as explained above, that the majority of fossils exposes the dorsal side. However, if the argument is that specimens being preserved with their ventral side up stand a lesser chance of preservation, this also makes sense: the ventral exoskeleton is considerably less sclerotised than the dorsal side. So, if a carcass is lying on the sea floor upside down, with its ventral side exposed, scavengers and currents will have free reign to tear it up. If, on the other hand, the carcass is not overturned, the heavy dorsal exoskeleton will to some extent help protect the more fragile ventral anatomy from being destroyed, allowing time for the carcass to be buried, potentially helping it to enter the fossil record. In this respect, a dorsal-side-up orientation may actually aid preservation of limbs and soft anatomy, rather than “damage” it, as the authors suggest. In that case, the overlying dorsal exoskeleton would obscure any underlying preserved parts, making it harder for them to be studied, but in itself would not “damage” them; in this respect, it is also important to note that at least in the case of non-biomineralising arthropods, flattening of carcasses mostly results from decay-induced collapse of the carcass onto itself, rather than from compaction by the surrounding sediment.

	On p. 19, lines 317-319, it is claimed that the Winterswijk Limulitella shows “extensive soft tissue traces”. At least based on the information provided in the paper by Zuber et al. (2017), this claim is entirely incorrect: the authors of that study only identified exoskeletal morphological features  of this fossil. They do refer to the presence of “muscular markings”, but as is clear from their figures, by this they do not mean actual muscle tissue, but rather the depressions on the prosomal shield associated with internal apodemes to which muscles would have attached. Hence, the only thing visible in their scans was the sclerotised, exoskeletal cuticle, which is neither soft, nor a tissue!

	Apart from these issues, I have added some further small remarks and (mainly typographical) corrections as sticky notes to the attached annotated manuscript.

	In conclusion, this manuscript does present some interesting data, and does merit to be published in PeerJ. However, before it can be published, the authors do need to rectify the several issues raised here.

Peter Van Roy
23 December 2021


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: AN EARLIEST TRIASSIC AGE FOR TASMANIOLIMULUS AND COMMENTS ON SYNCHROTRON TOMOGRAPHY OF GONDWANAN HORSESHOE CRABS
Review round: 2
Reviewer: 1

Output:
Basic reporting: I am fine with the changes that were made.
I only found one issue that needs to be checked: The section "Comments on application of synchrotron tomography to the study of fossil xiphosurids" in the discussion is not entirely the same in the PDF and in the DOCX. In the DOCX, this header appears two times, and then parts of the text are different in the two files. The authors need to have a close look here again to make sure that they submit the correct final version.
Experimental design: none
Validity of the findings: none
Additional comments: none


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: AN EARLIEST TRIASSIC AGE FOR TASMANIOLIMULUS AND COMMENTS ON SYNCHROTRON TOMOGRAPHY OF GONDWANAN HORSESHOE CRABS
Review round: 2
Reviewer: 2

Output:
Basic reporting: No comment.
Experimental design: No comment.
Validity of the findings: No comment.
Additional comments: The authors have made a genuine effort to accommodate the queries, suggestions and criticisms of both reviewers, which is appreciated. As a result, this revised manuscript is considerably better than the original version. Nevertheless, there are still some remaining issues the authors need to address before the paper can be accepted for publication:

- Both the abstract and Results section claim that the SRXT images revealed novel anatomical details beyond what was already known. In truth, this is an exaggeration: no genuinely new morphological/anatomical information was revealed; the SRXT scans only served to further emphasize and document some morphological details that were already known previously. I therefore strongly suggest the authors tone down their statements about the novelty of the findings to reflect this - indeed, in their concluding remarks, the authors themselves admit that the current study did not reveal 'much novel anatomy, nor traces of soft tissues'!

- The introduction still refers to 'ventral preservation' of specimens. As I discussed at length in my earlier review of the original manuscript, this term is ambiguous at best, and the authors should rephrase this to make clear what they mean exactly.

- In several places, the name of the Anisian stage of the Triassic is rendered as 'Ansian', dropping the first 'i'. This should be checked and corrected throughout the manuscript.

- The main purpose of CT and SRXT scanning is to reveal internal features of a specimen, or show features hidden in the matrix. This study did not really accomplish either of those goals, but did result in high-resolution 3D surface models of the specimens, which is worthwhile in and by itself. However, 3D digitalization of specimens in this way can be accomplished more easily with e.g. laser scanning, or to some extent even with photogrammetry and RTI imaging (although the latter two techniques have considerably lower resolution, and tend to suffer from artefacts, in particular in highly reflective areas). Therefore, the authors may consider adding a couple of words about these techniques to their discussion of the 3D digitalization of specimens.

- In a couple of places, 'parvinospinoslis' should read 'parvinospinosus'. Check!

- Moveable notches: sloppy terminology! The notches are not moveable - they accommodate moveable spines (which are not preserved in the specimen).

- Use Konservat-Lagerstätte instead of just 'Lagerstätte'. Seilacher distinguished two main types of Lagerstätten: Konservat-Lagerstätten, which are sites with exceptional preservation, and Konzentrat-Lagerstätten, which are sites characterized by an abundance of fossil material. Therefore, it is important to distinguish between the two - using just Lagerstätte makes it unclear what type of deposit you are referring to.

In addition to these remarks, I have made some smaller comments and (mostly typographical) corrections on the attached manuscript.

In summary, this revised manuscript is a marked improvement of the original, and if the authors address the remaining issues flagged here, the paper should be acceptable for publication.

Peter Van Roy
02 March 2022


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: LNCRNA MALAT1 MEDIATES OSTEOGENIC DIFFERENTIATION OF BONE MESENCHYMAL STEM CELLS BY SPONGING MIR-129-5P
Review round: 1
Reviewer: 1

Output:
Basic reporting: The introduction is very succinct and does not provide sufficient background for a large audience. The authors should explain differentiation of BMSCs in more details and introduce the key molecular players. Please also introduce lncRNAs and miRNAs; what they are and what they do.
The overall flow can be also improved. Currently the manuscript resemble a bullet point list of results rather than a story and sometimes lacks logical progression. For example, it is not clear why authors focus on miR-129-5p or miR-17-5p (line 248). These miRNAs were not introduced in the background section. Is it because Malat1 has binding sites of miR-129-5p and miR-17-5p?
The authors often refer the reader to Figures without telling the reader what their interpretations are. For example, lines 263–265 read, “The dynamic expression of osteoblastic markers Runx2, ALP and OCN during osteogenesis are shown in Figures 7B-D.” What are these changes? Does expression of Runx2, ALP and OCN increase or decrease during osteogenesis? Are these changes concomitant with changes in Malat1 expression?
Figures should be re-arranged. There are currently 15 Figures, and some panels are not cited in a chronological order (e.g., Figure 10B is referred in the text before Figure 10A). Also, please consider using a different color palette to facilitate interpretation for color-blind readers.
Figures 8J and 10J: what are the round inserts? I suppose, they are pictures of cell plates. If this is the case, indicate which region of the plate is shown below at a higher magnification, Also, the scale bar is barely visible.
Figure 9A: Runx2 signal cannot be seen easily, even in the Control sample, while in Figure 11A the signal appears fine. Please enhance the contrast for more visibility.
Experimental design: Please provide the reader all key information in the Material and Methods or Results sections, so that the reader does not need to search for it. For example, lines 220–221 read, “lncRNA-miRNA pairs (including 28 lncRNAs, 50 miRNAs and 281 mRNAs) were obtained as previously described (8).” Also, explain non-common abbreviations. What are NUSE values (line 212)? One does not need to introduce an acronym for an expression used only once in the text.
It is unclear to me which groups are compared in the section 3.3. Line 217 mentions “the tumor and adjacent normal tissue”, but lines 240–241 state “We found that lncRNA MALAT1 had higher node degrees, suggesting a potential role for BMSCs among elder donors.” Are tumor and normal tissues compared, or are tissues from young and old donors compared?
Validity of the findings: The authors should quantify their Western Blots. I am not convinced that there is a reduction in Stat1 levels.
Avoid expressions such as “slightly reduced” (lines 278 and 289) and “the change in luciferase activity was not obvious” (line 312). Please quantify your results and provide statistics.
The authors report that Malat1 is localized in the nucleus, therefore I am confused about its role as a miRNA sponge. miRNAs mature and function in the cytoplasm, so how does Malat1 interact with miRNAs? The authors perform AGO2 pull-downs and find miR-129-5p, Malat1 and Stat1 in the pull-down fraction. Stat1 and miR-129-5p make sense but finding nuclear Malat1 associated with AGO2 is surprising. I believe that this is an artefact and miR-129-5p binds Malat1 after the cells are lyzed and thereby compartmentalization is abolished.
It is not clear to me how Stat1 luciferase assay was performed. Did the authors clone miR-129 binding sites from Stat1 CDS into Luciferase 3’ UTR? If this is the case, I do not think that it is a meaningful experiment as even if miR-129 binds these sites, this binding is likely abolished by ribosomes upon translation.
Additional comments: The authors performed a lot of work and carried out an overall thorough analysis. I believe that points that I mentioned above will improve the 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: LNCRNA MALAT1 MEDIATES OSTEOGENIC DIFFERENTIATION OF BONE MESENCHYMAL STEM CELLS BY SPONGING MIR-129-5P
Review round: 1
Reviewer: 2

Output:
Basic reporting: The topic and content discussed in this manuscript are within the scope of the journal.

The English of your manuscript must be improved before resubmission. I strongly suggest that you obtain assistance from a colleague who is well-versed in English or whose native language is English.
Experimental design: The organization and subsections are also appropriate. The manuscript is structured and presented in a reader-friendly manner.
The survey methodology is consistent with comprehensive and unbiased coverage of the subject.
Validity of the findings: In this manuscript, there are well-developed and supported arguments that meet the goals set out in the Introduction, and the authors also provided very good descriptions and summaries of the important role of long noncoding RNA (lncRNA) metastasis-associated lung adenocarcinoma transcript 1 (Malat1). And the authors also indicated that lncRNA Malat1 may play a critical role in maintaining the osteoblast differentiation potential of BMSCs by sponging miR-129-5p.

The paper has minor, easily fixable, technical flaws that do not impact the validity of the main results
Additional comments: 1.	In line 217, I suggest using all Arabic numerals consistently
2.	Please indicate how many repeats are used in each experiment.
3.	In Figure 6B, the authors show Malat1 expression using qRT-PCR, I wonder if you use the internal control shown in Figure 7. I know that GAPDH amplification is not available in Nucleus. So, I don't know how the author got the result. I suggest the authors verify the expression of Malat1 by Western blot.
4.	In Figure 7, consider validating the expression of these genes at the protein level.
5.	In Figure 8A, I suggest the authors check knockdown Malat1 expression at the protein level. In Figure 8H, consider repeating 3 times in each group, and the quantitative analysis data of western blot analysis is missing, which should be presented.


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: LNCRNA MALAT1 MEDIATES OSTEOGENIC DIFFERENTIATION OF BONE MESENCHYMAL STEM CELLS BY SPONGING MIR-129-5P
Review round: 1
Reviewer: 3

Output:
Basic reporting: The authors should do proofreading more carefully
Experimental design: Methods:

What is the gender for the donors?

The age and quantity of the mice in each group should be mentioned in 2.5

Results:

Why was the bioinformatic analysis done in middle aged and elderly donors but animal-based study conducted in young and old mice?

Were 64 DELs and 651 DEMs identified between middle aged and old donors or between the tumor and adjacent normal tissues as stated in line 217?

In figure 7, did the authors do any statistical analysis or did they forget to label p-value in all plots?

In figure 8, 10 and 14, other than the mRNA quantification for those BMSCs differentiation markers, the authors should also quantify their changes at protein level via western blot analysis.

In figure 8(j) and 10(j), the authors should label which pictures are culture in GM or OM and explain in the result part how these support their conclusion.

The authors should add a MiR129-5p inhibitor only group in figure 14 (A)-(D) and a mRNA expression analysis of Malat1. As suggested by the mRNA analysis in figure 10, the inhibition of MiR129-5p itself can promote expression of BMSCs differentiation and the data in (A)-(D) so far are not sufficient to indicate a reversal effect on ASO-Malat1.

In figure 14 (F), the authors should consider adding a ASO+OM group and redo this assay since it is obviously in conflict with what has been shown in the ALP staining of figure (E).

Please check all the labels in each picture and make sure they are consistent throughout the manuscript. For example, the MiR129-5p inhibitor was presented as "inhibitor" in figure 10 and "MiR 129 inhibitor" in figure 14 figure A-D and F but MiR129-5p inhibitor in figure E.
Validity of the findings: The validity of the findings in this study is largely limited by the small sample sizes in both bioinformatic (n=5, n=4) and animal study (n=3).
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: LNCRNA MALAT1 MEDIATES OSTEOGENIC DIFFERENTIATION OF BONE MESENCHYMAL STEM CELLS BY SPONGING MIR-129-5P
Review round: 2
Reviewer: 1

Output:
Basic reporting: The authors addressed all of my comments.
Experimental design: N/A
Validity of the findings: N/A
Additional comments: N/A


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

Input:
Title: LNCRNA MALAT1 MEDIATES OSTEOGENIC DIFFERENTIATION OF BONE MESENCHYMAL STEM CELLS BY SPONGING MIR-129-5P
Review round: 2
Reviewer: 2

Output:
Basic reporting: The organization and subsections are also appropriate.
Experimental design: The topic and content discussed in this manuscript are within the scope of the journal.
Validity of the findings: It is better if the quality of the western blotting picture is higher.
Additional comments: No


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: FOLIAR NUTRIENT CONCENTRATIONS OF SIX NORTHERN HARDWOOD SPECIES RESPONDED TO NITROGEN AND PHOSPHORUS FERTILIZATION BUT DID NOT PREDICT TREE GROWTH
Review round: 1
Reviewer: 1

Output:
Basic reporting: The MELNHE experiment is a great service to forest biogeochemistry, and this paper represents an interesting update from the ongoing study.  Context from the larger experiment is communicated, and these findings are unique and worthy of their own paper.  However, the framing of the data requires revision before publication.
Experimental design: The data are rigorously collected and reported.  I particularly appreciate the reporting of negative results (mycorrhizal type, stand age).  Nonetheless, the busy figures suggest a research question that could use more honing.  Why show the slope of foliar nutrient change for all treatment types combined?  If a key hypothesis is that N addition will stimulate foliar P (or vice versa), panels of the main text figures should contrast treatment types and combine the factors that were not statistically interesting (like every species except pin cherry).
Validity of the findings: The work of this manuscript is framed in part as using foliar nutrient response to fertilization as an alternative measure of nutrient limitation to productivity.  However, this manuscript is better viewed as a direct test of whether foliar nutrition changes CAN indicate nutrient limitation to productivity.  It is akin to a methods test where a direct measure of nutrient limitation (tree trunk growth) is carried out in tandem with the controversial method of foliar chemistry.  The authors are correct to consider the possibility that nutrient limitation was missed by their tree trunk measurements.  An unmeasured biomass pool might have been stimulated by N+P fertilization.  However, especially because tree trunks are a large share of NPP in temperate forests and are perhaps the most important pool for decadal-scale carbon modeling of forests, the authors should also consider the possibility that their findings suggest foliar nutrition is not a useful metric of limitation.
Additional comments: Thank you for your work doing this research and preparing this manuscript.  My biggest piece of feedback is to reformat the figures to address a research question.  Here are specific notes line by line.

Line 38
Unhealthy leaves were excluded from your study, and there are open questions about the usefulness of foliar nutrition for assessing nutrient limitation (One example: Fisher JB, Malhi Y, Torres IC, Metcalfe DB, van de Weg MJ, Meir P, Silva-Espejo JE, Huasco WH. 2013. Nutrient limitation in rainforests and cloud forests along a 3,000-m elevation gradient in the Peruvian Andes. Oecologia 172:889–902.)

Line 80-82
This idea was originally proposed in: Koerselman W, Meuleman AFM. 1996. The Vegetation N: P Ratio: a New Tool to Detect the Nature of Nutrient Limitation. J Appl Ecol 33:1441–1450.

Line 107-109
More careful distinction is needed between this “study” within the long-term experiment (foliar nutrient response), and the long-term experiment as a whole (MELNHE).  This sentence also seems out of place when MELNHE is introduced later on.

Line 109-111
Broad justification of the long-term experiment is not necessary.  The manuscript should focus on justifying the questions it addresses within the long-term experiment.

Line 117-121
I suggest you state research questions here and focus on answering them throughout the paper.  These are two research questions I'd propose, based on what I find compelling in your data.  1. Can foliar chemistry (or changes in foliar chemistry) predict nutrient limitation to productivity, as measured by tree trunk growth? 2. Does adding one nutrient stimulate, inhibit, or not affect the status of another nutrient in foliage?

Line 158-159
Did you perform a sensitivity analysis on the inclusion of these edge trees?

Line 167-170
How many trees were sampled from edges?

Line 176-194
I like that you are sharing so many details about your protocol for future workers, but it is a bit distracting from the main story.  Your main story is deserving of attention!  I think this should go in the supplemental.

Line 200-201
I think I’ve missed something.  Were pre-treatment values measured three times (2008,2009,2010) in every plot? Also, what about Ca, Mg, and K?

Line 229-230
Could this have been affected by your choice to designate the variables carrying stand age information as random effects?

Line 260-262
The conventional placement of P in the denominator can lead to overstating the relative role of P as compared to N.  Use of log values is one solution, see: "The misuse of ratios in ecological stoichiometry" by Isles (2020) in Ecology.

Line 288-298
Vertical profiles of Ca, Mg, and K could be quite different than vertical profiles of N and P.  Plants might have more reason or ability to concentrate N and P in the uppermost sunlit leaves you sampled. The tiny K+ ion, in particular, is highly susceptible to leaching in upper leaves but less so in shaded leaves because throughfall water is loaded with K by the time it reaches shaded leaves.  Vertical leaf trait profiles have been published for sugar maple (e.g. Coble & Cavaleri's 2017 paper in Tree Physiology) and probably for your other species as well.

Line 315-316
By N allowing greater use of P for growth, are you suggesting N addition mobilized P to wood?  roots?  reproduction? If so, you might be interested in: Sardans J, Pen ̃uelas J. 2014. Trees increase their P: N ratio with size: Phosphorus; the treasure nutrient. Glob Ecol Biogeogr 24:147–156.

Line 319-320
Was the degree of P limitation (i.e. magnitude of tree trunk growth response to P addition) proportional to pre-treatment foliar N:P (supporting Koerselman & Meuleman’s 1996 hypothesis), magnitude foliar nutrient response to P application (evidence for the utility of foliar chemistry in assessing limitation to productivity), or neither (evidence against the utility of foliar chemistry in assessing limitation to productivity) ?

Line 390-392
I appreciate this context about the importance of Ca at the site.  What are the roles of Mg and K?  If site specific data do not exist, discussion of their role in temperate forests or even plants in general would help readers understand the motivation for the measurements.


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: FOLIAR NUTRIENT CONCENTRATIONS OF SIX NORTHERN HARDWOOD SPECIES RESPONDED TO NITROGEN AND PHOSPHORUS FERTILIZATION BUT DID NOT PREDICT TREE GROWTH
Review round: 1
Reviewer: 2

Output:
Basic reporting: This manuscript describes a unique experiment evaluating fertilizer additions of N and P and the effects on foliar nutrients on tree species in a northern hardwood forest.   The paper is well prepared, the figures are relevant, and the literature used in the paper is up to date and provides excellent context.

My only questions are in the use of the term "foliar base cations" -- usually referring to Ca and Mg (and sometimes K), which while they act as base cations in the soil, in the foliage are generally without charge, and it would be better to simply say something like "foliar Ca and Mg" .

Second, the term "tolerant" is often used to mean "shade tolerant".   Because plants have other tolerances (or lack thereof) it is best to be specific at each usage.

Why is N:P the only ratio evaluated?  The rational/context for only N:P could be better described.
Experimental design: Very well designed, and the QA/QC description is very thorough.
Validity of the findings: I find the use of relative changes, expressed as percentages, potentially misleading.  I accept that the authors do not actually statistically compare them, but a small change in a small number could result in a percentage similar to a large change in a large number.   The data are available to the reader, so they can be evaluated, fortunately.

In the introduction, the objective statement  includes the statement "We anticipated that tree species, stand age and site quality would also influence foliar responses."  Yet there is little if any,  reference to site quality in the discussion and conclusions .   It would be good to revisit the hypotheses/objectives clearly.
Additional comments: Well done study and very well done 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: FOLIAR NUTRIENT CONCENTRATIONS OF SIX NORTHERN HARDWOOD SPECIES RESPONDED TO NITROGEN AND PHOSPHORUS FERTILIZATION BUT DID NOT PREDICT TREE GROWTH
Review round: 2
Reviewer: 1

Output:
Basic reporting: My major revisions were addressed, particularly in framing a stronger research question.  This is a rigorous paper and a great addition to the literature.  I just have a few very small comments.  I will not need to see the paper again.
Experimental design: no comment
Validity of the findings: Figure 6: Are any of these trends significant?  And if so, significantly different from each other?  I am particularly interested in whether N+P addition eliminated the growth advantage of leaves that had higher Ca and Mg levels at the start of the study...and how N+P treatment perhaps even made a high starting potassium value into a disadvantage in growth.

Line 138: Were the exact same cations measured in the Fang study?

Line 1322: This comment about white birch demonstrates to me why N+P and +N should be treated separately.  Someone interested in whether trees can leverage added N to get more P would not be convinced by findings that include N+P data.  Combining the data might be relevant for some other research questions but not the one discussed here.
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: NICKEL MINE SOIL IS A POTENTIAL SOURCE FOR SOYBEAN PLANT GROWTH PROMOTING AND HEAVY METAL TOLERANT RHIZOBIA
Review round: 1
Reviewer: 1

Output:
Basic reporting: Comments for Author,

In this paper, The  Author tries to explore the " Nickel mine soil is a potential source for soybean plant growth-promoting and heavy metal tolerant rhizobia". The paper is the scope of Peerj. However,
 I have some comments before accepting this paper.

Comments for Author,

These sentences need more related papers. to improve please support it. " Heavy metal
 contamination is a risk to food security, ecological environment, and even to human health
 through bioaccumulation in the food chain"  a- https://doi.org/10.1007/978-3-030-89984-4_19

This  below sentences make no sense;

In this study, through the cultivation experiment of soybean plant in nickel mine soil, we
aimed to find effective plant growth promoting and heavy metal resistant strains to facilitate
phytoremediation of heavy metal contaminated soil.

Please rewrite the above sentences.

Please give the coordinate of soil that the Authors collected.

Plant growth ability test section should be reduced. There are some confusion

Best Regards
Experimental design: The design of the experiment was acceptable.
Validity of the findings: The results are reasosable.
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: NICKEL MINE SOIL IS A POTENTIAL SOURCE FOR SOYBEAN PLANT GROWTH PROMOTING AND HEAVY METAL TOLERANT RHIZOBIA
Review round: 1
Reviewer: 2

Output:
Basic reporting: In the current study, authors have isolated plant growth promoting symbiotic strains from nickel mine soils with the idea of using such isolates for phytoremediation. The manuscript is well written with detailed description of the methods and proper interpretation of the results.

Few minor suggestions/corrections are detailed in the edited version of the manuscript. Please address them.
Experimental design: All the research questions are well defined and authors have conducted all the necessary experimentals to support their study.
Validity of the findings: The result are well represented and all the necessary raw files were provided.
However, it would be good to include the sequencing files as a word or text file in a zip format so that it is easily accessible.
Authors should include title and figure legends or footnotes for all the tables and figures wherever applicable along with the abbreviations.
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: NICKEL MINE SOIL IS A POTENTIAL SOURCE FOR SOYBEAN PLANT GROWTH PROMOTING AND HEAVY METAL TOLERANT RHIZOBIA
Review round: 2
Reviewer: 1

Output:
Basic reporting: The Author improved the paper carefully, Now the paper could be accepted for publication.

Best Regards
Experimental design: Well
Validity of the findings: Well
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: NICKEL MINE SOIL IS A POTENTIAL SOURCE FOR SOYBEAN PLANT GROWTH PROMOTING AND HEAVY METAL TOLERANT RHIZOBIA
Review round: 2
Reviewer: 2

Output:
Basic reporting: Authors have addressed all the comments and suggestions made in earlier review report. All the corrections are satisfactory and the manuscript can be accepted in its current form.
Experimental design: No comments
Validity of the findings: No comments
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: DNA BARCODING FOR BIODIVERSITY ASSESSMENT: CROATIAN STONEFLIES (INSECTA: PLECOPTERA)
Review round: 1
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: This manuscript generated a validated national reference DNA barcode library for stoneflies in Croatia, which can support the implementation of cost-efficient DNA-based identifications and assessments to ecological status. The study will contribute to knowledge about the distribution of species, genetic lineages within species and systematic and phylogenetic relationships. This article appears to use standard methods for analyzing and presenting the data. The scientifically sound and the outcomes are relatively well explained. Therefore, I recommend its acceptance for publication in PeerJ after minor revision:
1) Line 90: The references should be carefully check, delete D and I in “D Hlebec & I Sivec, 2021”. The same errors also existed in Line 224, Line 394, etc.
2) Line 108: Popijač, 2008; Popijač & Sivec, 2009a, should be spelt in italic.
3) Line 223: Delete the last dot of “MrBayes 3.2.7.”.
4) Line 245: change ‘Taeniopteryx sp.’ to ‘Taeniopteryx sp.’. The same errors also existed in Line 390, Line 524, etc. Please check carefully.
5) Line 356: In the sentence ‘distance threshold 3,6%’, it should be 3.6%.


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: DNA BARCODING FOR BIODIVERSITY ASSESSMENT: CROATIAN STONEFLIES (INSECTA: PLECOPTERA)
Review round: 1
Reviewer: 2

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The manuscript is an important faunistic and genetic contribution. The authors present a comprehensive and taxonomically DNA barcode database for Croatian stoneflies, with the sufficient datas and analysis.

Comments and proposed corrections:

• The language of the manuscript is in need of a general check of a native speaker. As being not a native speaker, I avoid of making linguistic comments or corrections.

• Regarding Fig. 2: Almost all data are consistent with the family status, but Capnidae is split into two parts by the Taeniopterygidae dataset (in Fig. 2: 48, 50, 51). I suggest the authors to discuss this situation further.

• "Morphological variability" was repeatedly mentioned in the manuscript, and the attachment (peerj-69224-Supplemental_File_S1) also includes multiple photos of stoneflies. What puzzles me is that these beautiful images don't seem to present any morphological variability mentioned, or even contain any figures of taxonomic diagnosis (male terminalia , etc.). In addition, in these hundreds of datas, one species can correspond to several genetic sequences. So I wonder, do these sequences severally come from, and match the different morphotypes in each species? Is there some correlation between the morphological variability and their corresponding genetic diversity? Perhaps the authors can discuss more in this regard.