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: IL-6 AND TNF-Α SALIVARY LEVELS ACCORDING TO THE PERIODONTAL STATUS IN PORTUGUESE PREGNANT WOMEN
Review round: 1
Reviewer: 1

Output:
Basic reporting: English used in the manuscript needs improvement. Long ambiguous sentences are used, which make it confusing to read.

Literature references are sufficient.

Professional article structure utilized.

The results of the study are not sufficient to support the hypotheses.
Experimental design: Research is within the aims and scope of PeerJ.

It is not clearly stated how the research question really fills the gap in the knowledge. The question that the authors are asking is whether changes in salivary il-6 and TNF alpha levels can be utilized for diagnosis of periodontal disease in pregnant women. Diagnosis of periodontal disease requires a periodontal probe and measurement of attachment loss around teeth. What the additional benefits are of a salivary test using inflammatory markers is not clearly stated. In the past several studies have looked at the salivary levels of inflammatory cytokines in patients with periodontal disease, yet to date there are no salivary tests available. This discrepancy needs to be clearly addressed in the manuscript. It is also not clear why the authors chose only these two cytokines.

The level of investigation is not rigorous enough. For example, it is not clear at what point during the pregnancy, the saliva samples were collected. At least two to three specific time point based saliva collection and analysis during the course of pregnancy would be ideal. Because within the healthy group there could be natural changes in the inflammatory markers during the course of pregnancy. Also the authors are analyzing only two cytokines. An array of inflammatory markers are needed for more rigorous investigation.

Methods are not described with detail. For example it is not clear if smokers were excluded from this study. Also the time point of saliva collection is not mentioned and the method of saliva collection is not explained in detail.
Validity of the findings: Although there might be a statistical difference in the levels of the cytokines between the healthy and periodontitis groups I am not sure if there is a clinical significance as the differences in cytokine levels are not even twice the normal levels.

Statistical analysis is done by ANOVA. For clinical studies, it would be ideal to perform ODDS Ratio and Relative Risk analyses for more robust statistical outcomes.

Conclusions stated do not support the original question based on the weak methodology and limited results.
Additional comments: Good study but lacks rigorous investigation. More data collection points needed. More inflammatory cytokine profiling needed. Statistical analysis should include Specificity, Sensitivity, Odds Ratio and Relative Risk. English in the manuscript needs professional editing.


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: IL-6 AND TNF-Α SALIVARY LEVELS ACCORDING TO THE PERIODONTAL STATUS IN PORTUGUESE PREGNANT WOMEN
Review round: 1
Reviewer: 2

Output:
Basic reporting: The study entitled “IL-6 and TNF-α salivary levels according to the periodontal status in Portuguese pregnant women” is well designed, and has a robust hypothesis introducing the pathogenesis of the association between pregnancy and periodontal disease.

It has been well known that interleukin 1-beta is also an important pro-inflammatory cytokine playing an important role in pregnancy, pregnancy outcomes or pathologies of pregnancy, and periodontal disease. Thus, the authors should also introduce that the role of IL-1 beta in periodontal disease and pregnancy. Just as, IL-1 beta is a precursor and cytokine for IL-6 and TNF-α.
Experimental design: It is a well designed, and the relationship between hypothesis and results is well discussed.
Because of the fact that this is a cross-sectional study, it should be stated as an important limitation of this study.
Validity of the findings: The longitudinally studies suggesting the poor pregnancy outcomes (such as preterm birth, low birth weight, and preterm and low birth weight, etc ) and periodontal disease in the same population will also be helpful to clarify the effect of periodontal disease on pregnancy complications or poor pregnancy outcomes.. These statements should be added to the section of conlusion of the text for further consideration.
Additional comments: 1. The study entitled “IL-6 and TNF-α salivary levels according to the periodontal status in Portuguese pregnant women” is well designed, and has a robust hypothesis introducing the pathogenesis of the association between pregnancy and periodontal disease.
2. It has been well known that interleukin 1-beta is also an important pro-inflammatory cytokine playing an important role in pregnancy, pregnancy outcomes or pathologies of pregnancy, and periodontal disease. Thus, the authors should also introduce that the role of IL-1 beta in periodontal disease and pregnancy. Just as, IL-1 beta is a precursor and cytokine for IL-6 and TNF-α.
3. It is a well designed, and the relationship between hypothesis and results is well discussed.
4. Because of the fact that this is a cross-sectional study, it should be stated as an important limitation of this study.
5. The longitudinally studies suggesting the poor pregnancy outcomes (such as preterm birth, low birth weight, and preterm and low birth weight, etc ) and periodontal disease in the same population will also be helpful to clarify the effect of periodontal disease on pregnancy complications or poor pregnancy outcomes.. These statements should be added to the section of conlusion of the text for further consideration.


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: BIOINFORMATIC ANALYSIS AND IDENTIFICATION OF POTENTIAL PROGNOSTIC MICRORNAS AND MRNAS IN THYROID CANCER
Review round: 1
Reviewer: 1

Output:
Basic reporting: The manuscript from Jianing Tang, and colleagues identified the differentially expressed microRNAs and mRNAs between cancer tissues and matched normal thyroid tissues by public microRNA and mRNA sequencing data. They also investigated the mechanisms of thyroid cancer by constructed protein-protein interaction (PPI) network of differentially expressed mRNAs combined with microRNA-mRNA interaction analysis. Overall, I think the authors supplied useful candidate genes and theoretical basis of mechanisms for thyroid cancer research.

I have several major concerns about this paper.

1.	I agree with specific gene changes are associated with cancer initiation, however, what is the relationship between specific gene changes and gene expression change. I mean, a specific gene changed means its expression level change or its protein production change. If I am not understanding wrong, the DE genes identified in this manuscript between cancer tissue and normal tissue should be the same gene at different expression level in different tissues. I think the introduction needs to elaborate what kind of gene change the authors are looking at and explain more about that.

2.	What is the quality of raw microRNAs and mRNAs data used in this paper? I suggest add this information as a supplemental table here.

Minor concerns:
1.	Paragraph transitions need to improve in the introduction part.
2.	For Gene ontology and pathway enrichment analysis, I am wondering why not use GSEA for trancriptome analysis?
3.	Are the pvalues in Table1 and Table2 adjust p value? If yes, please label “adjust P value” in the tables.
4.	The resolution of figures need to be improved.
Experimental design: no comment
Validity of the findings: no comment
Additional comments: Generally,  I like this work and the idea generated by the authors. The introduction part really needed to improved and more detailed.


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: BIOINFORMATIC ANALYSIS AND IDENTIFICATION OF POTENTIAL PROGNOSTIC MICRORNAS AND MRNAS IN THYROID CANCER
Review round: 1
Reviewer: 2

Output:
Basic reporting: In this study, the authors re-analyse TCGA data from thyroid cancer samples in order to find micro RNAs (miRNAs) and mRNAs that can be used as prognostic factors. In order to do this, they do analyses to find differentially expressed miRNAs and mRNAs between cancer and normal samples, and then do GO-term enrichment analyses and build a protein-protein interaction network in order to try and devise biological pathways that might be altered in thyroid cancer. Generally, the Methods section needs substantially more detail to be able to follow what the authors have done, and to put the relevance of their results into context. This could be helped in the introduction by adding some references to previous studies that explore this same issue, and saying how this study differs from these previous ones and ideally how it improves the analysis. I think that figures and tables are well presented, even if hard to understand in a few cases (details below). The discussion needs to include not only results that agree with the authors’ analyses but also those that don’t, and it should be discussed or speculated why this may be the case. The manuscript is written in understandable English, but it can be improved (one example would be changing “…substantially promote prognosis” (lines 37-38) for “substantially improve prognosis”. Ideally, a replication of these results should be attempted if another dataset is available for analysis (maybe in dbGaP?).
Experimental design: In general, the Methods section needs much more detail to be understood and replicated.

In the Data Processing section, it is only mentioned that “clinical information of thyroid and RNA sequencing data from TCGA database” were obtained. No details are offered regarding the filters applied for downloading data, if any (for example, I assume they are talking about TCGA-THCA, thyroid carcinoma? Because there is also some thyroid data in the FM-AD project. Additionally, were all tissue types included? Did they download HTSeq raw counts, FPKM or FPKM-UQ data? Did they download only tumours with matched normal? In the second paragraph of this section authors mention that clinical information for patients included race: White, Black or Asian but there is one patient in the TCGA-THCA dataset with American Indian/Alaska Native ancestry. Was this patient excluded?). Authors also do not give any details about what they used to normalise these data, they just mention it was done “with R language package”. The package used and the initial dataset downloaded can have a huge impact on the results, for example, there are some R packages that already perform normalisation and thus raw HTSeq counts must be used with these. Clarifying these details is necessary before a reader can understand the methods.

In the Kaplan-Meier/log-rank analyses, it is mentioned that a p-value threshold of 0.05 was used to determine the prognostic value of differentially expressed miRNAs, but it is not mentioned if a multiple testing correction was applied to these p-values.

In the Gene Ontology and pathway enrichment analysis, what methods were used to find enriched GO terms? It is mentioned that online tools from the website DAVID were used, but it is not mentioned which ones. What version of the tools were used? Does it use hypergeometric tests to obtain enriched pathways? What identifiers were used?

The same with the protein-protein interaction network: What version of STRING was used? What of the many different datasets were downloaded? Also, what a “significant module of the PPI network” is needs to be clarified, and also what the software CytoHubba does.

Could the authors also please specify the versions of software used to predict the targets of miRNAs, and also, were all targets predicted for each gene used? Was some kind of cut-off value for confidence used?

In general, citations to all packages used need to be added, as at the moment only some of these have a reference.
Validity of the findings: Given that there are many details missing from the Methods section, it is difficult to establish how sound their results are. Specific concerns:

-	It is mentioned that 514 thyroid cancer samples were analysed, but only 59 matched normals. How were the differentially expressed genes obtained then? I would think that only the tumour tissues with matched normal could be used for such comparison, but 514 tumour samples seems excessive. Can the authors clarify this point please?

-	It is mentioned that 7/72 miRNAs were related to overall survival. How likely is it that this result is purely due to chance? Looking at the Kaplan-Meier survival curves, these look very similar (for example, b and f could be the same graph, at least by eye). It seems to me that it is the same two groups of patients (or two very similar groups) in all graphs. It could very well be that any these 7 miRNAs distinguishes these two groups of patients (although their effects are very possibly highly correlated), but it would be good if this analysis could be replicated in another dataset – is there any other available where this could be done? Reviewing the literature, it does seem that miR-146b has also been found upregulated in thyroid tumours vs normal (though only in PTC), but I have not found evidence for the other miRNAs. There are other studies exploring this issue, which the authors do mention in the Discussion – but only mentioning those results that agree with theirs. Could the authors add a note speculating why their results might be different from these, especially as it seems that miR-221 and miR-222 have been described previously but do not come up in the authors’ analyses?

-	It is stated that 6 target genes regulated by 3 miRNAs were found in a list of 1,766 genes. How likely is it to find this result by chance? If the authors perform a randomisation experiment in which they iteratively select 1,766 genes from the genome, how often do they find 6 or more target genes of the 7 chosen miRNAs?

-	It is difficult for me to understand whether the gene ontology analyses mean anything at all. It seems like a long list of processes and pathways – nearly any list inputted into DAVID will give a result. How is this result meaningful?

-	Figure 3 is hard to interpret – all comparisons showed there seem to be significant (they have an asterisk above the two groups in all cases).  Are they only showing the significant comparisons among all possible comparisons? And if so, did they correct for multitesting? What test was used to compare the groups? The effect sizes seem to be vanishingly small - Significance may arise from the number of individuals in each group but effects may not be meaningful in a clinical setting. Could the authors discuss a bit about the utility of this result as a prognostic factor?
Additional comments: In general and to summarise, I believe the most important points to address before this study can be accepted for publication are:

-	Addition of detailed description of Methods
-	Perform randomizations / multiple testing corrections where needed to control for false positives
-	Discuss results that agree with the authors’ results, as well as those that don’t


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: BIOINFORMATIC ANALYSIS AND IDENTIFICATION OF POTENTIAL PROGNOSTIC MICRORNAS AND MRNAS IN THYROID CANCER
Review round: 2
Reviewer: 1

Output:
Basic reporting: This manuscript does a good job on revision. I am happy with the improved figures and languages. This work supply useful candidate genes and theoretical basis of mechanisms for thyroid cancer research.
Experimental design: I was pleased to see more details were added to the methods part. The experiment was well designed.
Validity of the findings: No comment.
Additional comments: Overall, the authors did a good job on revision. All my questions were well addressed. I agree to accept this manuscript.


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

Input:
Title: BIOINFORMATIC ANALYSIS AND IDENTIFICATION OF POTENTIAL PROGNOSTIC MICRORNAS AND MRNAS IN THYROID CANCER
Review round: 2
Reviewer: 2

Output:
Basic reporting: The authors have added substantially more detail to their methods, I am thankful for that as it is much easier to follow the performed analyses now. They have also added more references, discussed previous results, and added a couple of figures to clarify their methods.

I do believe the study is much more complete now and is suitable for publication.
Experimental design: The research question is well defined. I believe that the authors have followed logical steps to reach their conclusions. Methods now have sufficient detail for replication.

Some minor comments:

Can a sentence clarifying that a network interaction analysis was performed be added to the abstract, before talking about the top 10 hub genes? That would help make the abstract more understandable.

In line 148, should it say FDR < 0.05?

In line 174, would it be more understandable if it were worded as “A total of 573 samples with microRNA sequencing data and 573 samples with mRNA sequencing data were analyzed….”?

In line 176, it says that “RNA sequencing data included 510 thyroid cancer samples and 58 matched normal samples”, but this sums to 568 whereas the total number of samples analysed is stated as 573. What are the other 5?
Validity of the findings: This comment may stem from my limited experience with Kaplan-Meier survival curves, but I would suspect that the miRNAs and LPAR5 mRNA that are associated with survival are prognostic factors but not completely independent from each other: e.g. the survival curves for each of these look very similar. However, if these have been all evaluated together (i.e. each of these RNAs has an effect even after controlling for all others), then I am happy with the conclusions.

For this revision, the authors performed a replication of their results with additional available datasets, a very welcome addition to their study. This additional analysis has a large overlap with their own results, which lends credibility to their methods and conclusion.
Additional comments: I thank the authors for their answers and for adding details to their text and performing the additional suggested analyses.


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: POST-TRAUMATIC STRESS SYMPTOMS ARE ASSOCIATED WITH BETTER PERFORMANCE ON A DELAYED MATCH-TO-POSITION TASK
Review round: 1
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The current study examined the association of PTSD with performance in spatial and temporal discrimination task. Three groups of participants were recruited, civilian, non-combat veteran, and combat veteran, based on their symptom severity. The key findings were that individuals with PTSS had higher accuracy rate in the spatial discrimination task without compromised prolonged RT. To further explore the possible accounts to the unexpected performance of individuals with PTSS, the authors conducted a second experiment using the Drift Diffusion Model (DDM), and realized that individuals with PTSS could accumulate evidence faster and have lower response caution during decision making. Generally speaking, the current manuscript is well-written and can provide insightful information to the decision-making process of PTSD. However, several caveats should be taken care of by the authors before being published on the PeeJ.

Introduction-
1.	The authors are recommended to reorganize the paragraphs regarding behavioral and neurobiological correlates of PTSD and non-PTSD. Maybe they can address the behavioral findings first (e.g., findings in associative learning task), and then go to the neurobiological aspects (i.e., findings in hippocampus) in another paragraph. Currently, the authors address this topic interchangeably between behavioral and neurobiological findings, which make the flow difficult to follow.

Method-
2.	The major shortcoming of the current manuscript is the use of non-clinical assessment for PTSD. The authors are strongly recommended to justify the use of these self-assessed checklist and scale considering that their findings are basically inconsistent with that from others.
3.	The authors are suggested to justify the examination for cognitive flexibility as well as the use of the TMT.
4.	The description of the spatial discrimination task should be consolidated. The current version is somewhat lengthy.
5.	Also, the authors should provide additional information for why they included “gender” as a factor into data analysis. Are they seeking for a gender-moderating effect?

Experiment 1 & 2 Discussions-
6.	Discussions for experiment 1 & 2 should be consolidated to shorter length, and leave most of the major discussions to the General Discussion.

General Discussion-
7.	See 6.
8.	Generally, the discussion are easy-to-follow except that the authors are suggested to provide some explanation/mechanism to why the PTSS group outperformed the non-PTSS group

Minor issues-
9.	The scheme of the spatial discrimination task in Figure 1 should be revised. The graphic in the current version is not clear enough.


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: POST-TRAUMATIC STRESS SYMPTOMS ARE ASSOCIATED WITH BETTER PERFORMANCE ON A DELAYED MATCH-TO-POSITION TASK
Review round: 1
Reviewer: 2

Output:
Basic reporting: Authors of the current study examined visual discrimination using a pattern separation task in subjects with PTSD symptoms (PTSS). Subjects with PTSS showed better discrimination than matched healthy controls which was not linked to slower response time. The authors argue that this links to previously reported PTSD-related overgeneralization associated with hippocampal abnormalities. Finally, the authors used a diffusion model to examine the task accuracy and RT. They found a faster rate of evidence accumulation and lower threshold for the PTSS group.

Based on previous work by the authors, subjects with PTSS showed enhanced generalization with higher levels of PTSD symptoms (re-experiencing). In the current study, however, the authors present results in the exact opposite direction where subjects with PTSS are better at differentiation (discrimination), which is the opposite of compression (generalization). It would be very useful if the authors can provide a theoretical framework to mend the gap between these different empirical findings.  For example, the author's aforementioned work on generalization in PTSS is not sufficiently discussed in the introduction or discussion to explain the difference between impaired generalization, decreased generalization, increased generalization, and over-generalization.

Finally, the authors ought to consider the various studies on visual discrimination in human and animal models of PTSD that can significantly improve the current arguments.
Experimental design: There could be an important confound in the task if the Simon effect is controlled for where learning is better for congruent (on the same side) rather than noncongruent stimulus-response associations.

Did the authors examine correlations between questionnaires and task results to look for potential covariates to include the ANOVA model?

The authors report exclusion of various trials with long or short RTs. What was the adopted criterion for this exclusion?

In the modeling section, how was the model fits performed? Which parameters were varied according to trial type and which ones were held constant? A bit more detail in this section would help the reader understand the modeling approach better.
Validity of the findings: Although the findings present an interesting new direction in this field of research, the current results still require further investigation to clarify the variables in action. The results are robust and the analytical approaches are sound. It would be very useful if the authors can add a measure of effect size.

The main issue with the current study is linking the current findings with previous literature on PTSD and generalization. Further, there is a wealth of human and animal literature on pattern separation and the hippocampus that can improve to the introduction/discussion sections.
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: POST-TRAUMATIC STRESS SYMPTOMS ARE ASSOCIATED WITH BETTER PERFORMANCE ON A DELAYED MATCH-TO-POSITION TASK
Review round: 2
Reviewer: 1

Output:
Basic reporting: Minor issue:
Line 43- should be "respond differently" rather than "respond different"?

Data analysis-
For clarification, please add texts elaborating the statistics for grouop homogeneity (chi-square test and t-tests), as shown in Table 1.
Also, though the authors did not found group difference in demographic measures such as gender ratio or education, texts elaborating control for these factors if group homogeneity was not met (e.g., descriptions to ANCOVA) are needed.
Experimental design: N/A
Validity of the findings: Just come to mind, there should be some factors which may affect cognitive function (e.g., socioeconomic status, intelligence) but were not adjusted for in the current study. This issue should be briefly discussed. Likewise, there may be comorbidity relating to PTSD (e.g., depressive symptoms) which were not controlled for in the current study. The authors should consider these limitations and elaborate how these limitations can be mitigated in the current study.
Additional comments: The authors did a good job in revising the manuscript. I am satisfied with most of the modifications made to the manuscript except several minor issue left to the authors for consideration.


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: POST-TRAUMATIC STRESS SYMPTOMS ARE ASSOCIATED WITH BETTER PERFORMANCE ON A DELAYED MATCH-TO-POSITION TASK
Review round: 2
Reviewer: 2

Output:
Basic reporting: No further comments.
Experimental design: No further comments.
Validity of the findings: No further comments.
Additional comments: The authors addressed all points raised initially.  I have no further comments.  This is a very interesting paper that will present a valuable addition to the PTSD literature.


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: PATTERNS OF GENETIC STRUCTURING AT THE NORTHERN LIMITS OF THE AUSTRALIAN SMELT (RETROPINNA SEMONI) CRYPTIC SPECIES COMPLEX
Review round: 1
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: Islam et al. address population genetic structure of a northern group of the Australian smelt. They found high levels of genetic differentiation both among and within rivers, suggesting limited connectivity among populations.
	Although the scientific novelty of this study is limited, their findings would be informative to the local resource managers. They hired standard population genetic procedures to find clear genetic differentiations among populations, and their conclusion seems to be reasonable. Their writing is clear in general.

Specific comments

Ln. 38: Is the comma necessary?

Ln. 85-104: I don’t find a strong connection between the theme of the manuscript and the species diversity in Australia argued here.

Ln. 113: What about the faithfulness of the homing behavior in this species? Given that even a small amount of reproduction by straying fish can homogenize genetic compositions between populations, it is meaningful to inform readers how much is known about the stray rates of this and the related species.

Ln. 130: Italic for gene name? (throughout the manuscript)

Ln. 179: fifteen or 15? Be consistent throughout the manuscript.

Ln. 247: Use citations for acknowledging the model and the method, even though they are implemented in a software.

Ln 258: Share which populations/loci exhibited deviations from HWE with readers, so as the instances of null alleles and significant LDs.

Ln. 283: What does ‘initially’ mean here? The authors argue that the highest likelihood at K=8 in Ln. 289.

Ln. 323: Provide accession numbers before the acceptance of the manuscript.

Table 1: Inclusion of population samples with n=8 in population genetic structure analyses is misleading. Try with and without these population samples and argue if their conclusions are supported even without these samples. It may also be reasonable to pool upper-lower population samples for these two cases, given the low Fst values within each river.

Table 5: What if P<0.05 was employed for relaxing the detection criteria? Sharing the data is ideal, and an argument on the selection criteria in the manuscript is a must.

Fig. 5: Use different colours for Mary and Coomera. Light yellow characters for Logan/Albert are hard to read.


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: PATTERNS OF GENETIC STRUCTURING AT THE NORTHERN LIMITS OF THE AUSTRALIAN SMELT (RETROPINNA SEMONI) CRYPTIC SPECIES COMPLEX
Review round: 1
Reviewer: 2

Output:
Basic reporting: Some of the punctuation and grammar was sub-optimal, leaving the reader confused about the points being made. There is inconsistent formatting of large numbers, inappropriate CAPS in several instances and scientific names that are not italicized.

I have provided commentary in my review attachment on instances where it needs to be improved.

I found parts of the introduction and discussion to be redundant. The authors may feel they were providing context to their study. But I consider the context provided didn't really hit the mark of explaining what it was that their study was trying to achieve.

There are some basic errors in simple parameters such as the family name of the study species, latitude swapped with longitude in Table 1 and an error in the tally of mtDNA samples processed. If the authors erroneously reported these data, how can the reader trust the accuracy of the more complicated figures?

The Genbank accession numbers where not provided in the manuscript. Only place-filling XXXXXX-XXXXX was provided.

The captions to the tables and figures were generally not sufficiently detailed to describe what was presented.

The resolution of figure 3 needs to be improved.

Suggestions for improvement are provided in my attached review.
Experimental design: Semi-original research that confirmed pre-existing knowledge on population structure of the relevant populations.

The authors clearly stated their hypothesis and posed the question of population structure relevant to life history strategies in previously un-studied northern populations of their study species (which was new). But they didn't really explain why this information is necessary or important.

The analyses used were rigorous. However, the authors could have made greater attempts to achieve their target sample sizes per population (this is an abundant species that is easy to catch) and they could have scrutinised the outputs of their analyses more thoroughly given what looks like one or two aberrent individuals in one population which are likely to have substantially contributed towards the results reported.

The methods were described in sufficient detail to be replicated.
Validity of the findings: Sample sizes were quite small in a few populations.

 One or two aberrent individuals in one population are likely to have substantially contributed towards the results observed. The data from this/these individuals should be scrutinised. If it is found to be of questionable origin, then this sample should be omitted from the data and the data re-analysed.

The conclusions based on the objectives and hypotheses could be emphasised a little more. At present, the conclusions focus on the definition of units of conservation and don't relate all that well to the theme of dispersal and life history strategies.
Additional comments: Line 26: Family is Retropinnidae, not Retropinninae.
Line 26: It can’t simply be defined as a facultative diadromous species? The next sentence suggests other proposed definitions of movement behaviour, of which facultative diadromy is only one.
Line 32: The hypothesis is fine, but seems at odds with the description of life history/movement behaviour suggested (facultative diadromous). It would be fine under the assumption that they are more inclined to potamodromy rather than diadromy. It is explained pretty well in the sentence starting on line 127. Could you rephrase the hypothesis in the context of the objectives?
Line 38: The sentence starting on this line should be shifted up to precede the previous sentence.
Line 40: Before making this claim, you need to demonstrate that there are no natural or artificial fish passage barriers between your within-river sampling sites.
Line 42: Delete ‘Overall,’
Line 42: Insert ‘Australian’ before ‘smelt’.
Line 42: Insert ‘comparatively’ before ‘higher’.
Line 45: Specify that the 8 MUs are within Queensland. There are certainly much more at a national scale.
Line 64: The sentence that starts on this line needs punctuation.
Line 65: Replace ‘reproduction’ with ‘reproducing’.
Lines 57-72: Is the 1st paragraph necessary? I don’t think it adds any value to the reader.
Line 82: You don’t need the word ‘genetically’ here.
Line 88: You don’t need the word ‘as’ here.
Lines 85-104: Is the 2nd paragraph necessary? I don’t think it adds any value to the reader.
Line 106: The genus should be called the Southern smelts given that Australian smelt is specific to R. semoni. Family is Retropinnidae, not Retropinninae. The sentence should read “Southern smelt (Retropinnidae: Retropinna) are abundant fishes distributed …..”
Line 109: Southern smelts are currently recognised …..
Line 126: Add comma after ‘individuals’.
Line 128: replace ‘of’ with ‘in’.
Lines 129-130: Sentence should start “Mitochondrial DNA sequence data combined with nuclear data .…”
Lines 131-133: These hypothesese are fine – albeit a little uninspiring. Why is this description of the hypotheses different from that in the abstract?
Line 139: Replace “each river except ….” with “each of eight river systems (except the Noosa River – downstream only).”
Line 140: Delete “Where possible,” and start sentence with We. It was always possible to catch more than 30 (as you aimed to do). You just didn’t manage to.
Line 147: Delete “All subsequent”. That can be inferred.
Line 148: PCR acronym used 1st here, but spelled out on line 151,
Line 152: Check the journals format for the o superscript in degrees celcius. You have used the number 0. They may want a lower case o.
Line 154: delete “amplified”. That can be inferred.
Lines 159-164: Why aren’t these samples shown in Table 1. Wouldn’t that be simpler. You could have a part a and part b for usat v mtDNA samples.
Line 164: (2 x 15) +10+10+9+8 = 67. Where did the extra sample come from?
Line 176: Replace ‘calculated’ with ‘undertaken’.
Line 180: ARLEQUIN v3.5.1.2 (Excoffier & Lischer, 2010) is cited in full here. Does it need to be cited in full in every subsequent usage (ie. line 194, 198, 225). Or just ARLEQUIN as on line 190?
Line 182: Is ARpriv equivalent to what is labelled PAR in table 1? If so, use a consistent symbology.
Line 183: Replace ‘estimated’ with calculated’.
Line 191: Replace ‘by’ with ‘through’.
Line 191: You have used a comma as a 1,000s place marker in this large number 10,000 (and others). But you have not in several other places (ie. line 196, 224, 236, 248 etc). Be consistent.
Line 201: Add “eight groups,” immediately after b).
Line 202: It is unclear what “according to the connectivity of streams to the upper river reaches” means.
Line 225: Arlequin is in lower case here. Should it be capitalised?
Line 231: Add comma after ‘generations’.
Line 238: End sentence at “… 2003).” Restart next sentence at “To estimate …”
Line 239: Was the analysis singular or should 3 repeat analyses be plural?
Line 239: Is “different random number of seeds” accurate wording? Do you use a number of seeds? Or several different seed numbers?
Line 243: Add “the” before “Paetkau”
Line 247: Lower case N in neighbour-joining.
Line 251: if JN232588 lacks locality information, why did you use it. You haven’t justified your speculation that it belongs to a southern lineage. Why not ask Chris Burridge?
Line 253: Shouldn’t the reference be to Hughes et al. 2014 rather than 2015?
 Line 267 and 269: What is the difference between Mean number of alleles and allelic richness averaged across loci?
Line 269-270: Why was it necessary to standardize sample sizes across populations at 6 individuals. Please explain.
Line 283: Be careful with the word “initially”. Did STRUCTURE determine this? Later it appears that STRUCTURE revealed the highest likelihood of 8 clusters. Both sentences can’t be correct. It is very confusing.
Line 285-287: It is interesting that the STRUCTURE outputs are counter to the FST results. Based on FST you would have expected the CEQ pops to separate in STRUCTURE before the SEQ populations. Why did this happen? Can the authors speculate?
Line 286-287: The second half of this sentence needs to be reworded. It is unclear.
Line 298-300: The sentence spanning these lines is very confusing – switching between singular and plural. Pleas reword. And define what ‘they”are/is.
Line 300-301: Again – counter to what happened in STRUCTURE.
Line 301: Replace “in” with “within”
Line 315: Capitalise C in Creek and replace “was” with “were”.
Line 323: GenBank accession numbers are “XXXXXX-XXXXXXX”.
Line 325-327: The result is consistent with STRUCTURE result rather than FST.
Line 334-335. Delete “using mtDNA and microsatellites”. It can be inferred.
Line 343: Delete “ of R. semoni”. It can be inferred.
Line 347-348: Was Tinana Creek (MRD) always differentiated because of the influence of the one (or perhaps 2) distinctly different individual(s) within the sample. Could this individual represent a mis-labelling error? What happens to the results if this individual is omitted from all the analyses? This is a serious issue that needs to be checked. Why didn't the BAYESASS and GENECLASS 2 analyses pull out these individuals as migrants when STRUCTURE clearly assigns them as such?
Line 349-350. Why is the reference to Hughes et al. (2015) relevant here?
Line 353: Lower case m in mariensis.
Line 355: Cherax disper = Cherax dispar. Is lung fish = lungfish?
Line 358-374: I found this paragraph lacked focus. It is hard to determine what point you are actually trying to get across. It starts OK = CEQ more highly structured than SEQ. But none of the subsequent sentences actually address issues relevant to why CEQ and SEQ differed.
Line 358: So why did STRUCTURE pull out the Brisbane River from within the SEQ populations before splitting any of the CEQ populations?
Line 366: Please define ’eustasy’.
Line 376: An alternative model of what? Please define.
Line 407: Replace “relative to lineages of R. semoni from the south of this species range” with “relative to more southern lineages of R. semoni”.
Line 415: Is the 10-60 km figure linear distance (crow flies) or river distance or between estuary mouths?
Line 417: “this species appears to occur in pools” is inaccurate speculation. They occur in all freshwater habitats. In fact, they are often most abundant in run and riffle habitats relative to pool habitats.
Lines 418-421. Evidence of migration within rivers does not constitute evidence of potamodromous migration. Dispersal within a river system can occur without it being a migration – which requires a regular population-level movement for a specific purpose.
Line 422: Reword – “where large number of Australian smelt was found…”
Line 421: Replace ”suggesting” with “suggests”.
 Line 424-427: This sentence needs to be reworded.
Line 430-447: While these statements are appropriate, they are too off-topic to constitute a conclusions section in the paper.
Line 433: Replace “Australian” with “Southern”.
Line 435: Delete “names”
Line 438: Delete “recognized that are geographically”.
Line 439: Delete “ these lineages”.
Line 443: Reword “where conservation strategy should be specified accurately”
Line 444: You haven’t established that anyone is actually proposing any translocations. Add context or delete.
Line 449: Replace “Alternatively” with “Further”. ESUs and Mus are not mutually exclusive. And specify that there 8 MUs in Queensland. There will be many more nationally.
Line 452: catchment should be plural.
Line 464: Mogurnda should be italicised.
Line 479: Volume should be bold font.
Line 509: Check that Crook used Retropinninae rather than Retropinnidae.
Line 524: Indentifying mis-spelled.
Line 549: Lower case m in mariensis.
Line 597: Add a space between genus and name of Craterocephalus stercusmuscarum.
Line 601: Italicise Hypseleotris compressa.
Line 620: Volume number should be bold font.
Line 634: Volume number should be bold font.
Line 669:  Italicise Hypseleotris.
Line 674: Volume number should be bold font.
Figure and Table captions are all generally insufficiently detailed and should be expanded upon.
Figure 1: This is not a map of Queensland. It is a map of south east Queensland spanning the Gold Coast to southern Fraser Coast. Major rivers and towns need to be added.
 Figure 3: The resolution of the figure is poor.
Table 1: Lower case s in sample. Neither PAR or Population specific FST are mentioned in the caption. Perhaps specify that these statistics are for microsatellite loci. The latitude and longitude values do not coincide with the column headings. Switch them.
Table 2: Capitalise B in Bonferroni.
Table 3: Define FST, FCT and FSC in the caption.
Table 4: One what basis are the most relevant results most relevant?
Table 5: migrant = migrants. End sentence after “…. location.”. Restart next sentence at “Results ….”.


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: PATTERNS OF GENETIC STRUCTURING AT THE NORTHERN LIMITS OF THE AUSTRALIAN SMELT (RETROPINNA SEMONI) CRYPTIC SPECIES COMPLEX
Review round: 2
Reviewer: 1

Output:
Basic reporting: I think it is ready for publication now, with one minor correction in Ln. 196 (‘micosatellite’ -> ‘microsatellite’).
Experimental design: ok
Validity of the findings: acceptable
Additional comments: I think it is ready for publication now, with one minor correction in Ln. 196 (‘micosatellite’ -> ‘microsatellite’).


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: STINGING WASPS (HYMENOPTERA: ACULEATA), WHICH SPECIES HAVE THE LONGEST STING?
Review round: 1
Reviewer: 1

Output:
Basic reporting: The basic reporting is sound.  A few more details are presented in the comments to authors panel.
Experimental design: The experimental design seems sound.  A few minor comments are also listed in the comments to authors section below.
Validity of the findings: I find the validity of findings sound with the minor suggestion that the discussion on host biology is a bit weak and is really unnecessary.  Its omission would not hurt the manuscript overall, rather might improve it.
Additional comments: The manuscript addresses some interesting questions relating to the importance of the aculeate hymenopteran sting length and various venom and defensive aspects of the insects.  The presentation is mostly straightforward and readily understandable.

In the Methods section, the description of how the sting length is measured could be improved.  The authors did not explain in detail how the length was determined; that is, was the length taken as the shortest distance between the triangular plate and the tip of the sting, or was some adjustment for the curvature and the length of the curve part of the sting measured?  Many of the stings are curved, sometimes strongly curved as in the Mutillidae, and this length seems more suitable to use rather than simply the shortest route from one point to the next.  When the sting is used it follows the shape of the sting shaft not the shape of the shortest distance between the triangular plate and the tip.

In the Results Table 1 only two ants are represented, yet ants are a crucially important group that are often highly toxic and painful and the study would benefit greatly by addition of a few more species – for example a Pogonomyrmex, an Odontomachus, and a Neoponera.  These are all common species that should be readily available and would add important information to the study.  This might also add more data to strengthen the evaluation of sting length versus venom painfulness and insect sociality.

Also for Table 1 the order would make more sense to have the taxa listed according to phylogeneny as presented in Branstetter et al 2017.  This would help the reader visualize the importance of phylogeny versus biology and lifestyle.

Figure 1b would be improved with two changes: first, the axes need units; and second, since the Pompilidae are represented by numerous species and are an important comparison to the Mutillidae, I recommend making another line of a different color for them just as was done for the Mutillidae.

Figure 2 could be strengthened if how the points were obtained was clarified; that is, is each point the average for an entire family, or for an individual species?  This might be important as some families have many representative taxa and others have only one.

I suggest the presentation relating to hosts and relative sting length be eliminated.  This is because the many points from the Mutillidae, all of whom parasitize Hymenoptera, bias the overall relationship.  Ideally we would compare equal numbers of taxa in each host or dietary type.  I do not think eliminating this information would overall weaken the manuscript, rather would strengthen it.

Some of the references are missing journal volume and page numbers.  These presumably will be added in the final version.


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

Input:
Title: STINGING WASPS (HYMENOPTERA: ACULEATA), WHICH SPECIES HAVE THE LONGEST STING?
Review round: 1
Reviewer: 2

Output:
Basic reporting: This manuscript presents data on sting length in the aculeate Hymenoptera, and interspecifically compares sting length to other aspects of wasp behavior and ecology to better understand sting length evolution.  I think this is an interesting topic on an an interesting group of organisms.  The paper is easy to read, and the figures are also easy to read and visually appealing.  The literature is well-discussed.  I have two major comments in this category:

1.  The discussion is devoted to presenting interesting adaptive hypotheses explaining the long stings of velvet ants.  However, it’s typical that the main results presented in the results section and figures are at least mentioned in the discussion.  It seems that the discussion addresses the title question: Which species have the longest sting?, but overlooks the other questions asked with your comparative analyses.  I would add a paragraph or two to discuss the relationship between (relative) sting length and toxicity, and the lack of a relationship between sting length and pain.  When doing so, it would be important to point out that the pain estimates are likely rather noisy, given the method used.  These patterns are mentioned in the abstract and the results section, and thus should be brought up in the discussion as well.

2.  I was able to open and see the data and code in the supplemental files.  However, I think it would be helpful to include an additional file explaining the columns in each file.  For example, I don’t know what the column “kind” with values 1000 or 10 refers to.  And why does each entry have a “velvet sting” and a “wasp sting” category?  Unless I’m misunderstanding the dataset, each line in the sting_data.txt is a measured sample.  If so, please include the species name, or further clarify what these data are.
Experimental design: I think the experimental design is OK, but has two drawbacks.

1.  L134: The method of inferring pain scores seems prone to bias and noise. Were you going strictly by the nearest phylogenetic relative with an independent pain report?  How many of the samples were “imputed” in this way?  Given that species can vary in pain rather a lot even within genera (e.g. Pogonomyrmex), I think inferring pain levels from different FAMILIES is likely to be rather noisy.  Pointing out that these data are likely to be noisy, and that this could influence the negative result you found, seems important.

2.  Statistical power in phylogenetic comparisons
The authors report an association between sting length and host, as well as defensive behavior (last line of the abstract, and throughout the discussion).  I think the adaptive hypotheses presented in the discussion (host use and defense) are good candidates to explain the longer stings of the velvet ants.  However, it is also important to note that the sting length association with host type (and perhaps with defensive behaviors) is the result of a single phylogenetic origin in the velvet ants.  Thus, the statistical evidence for an association with host type is less strong than assumed here.  This doesn’t mean that the suggested hypotheses are wrong, but this limitation should be pointed out.  In principle, any shared, derived feature of that group is as statistically associated with long sting length as host type.  I don’t think it is necessary for this manuscript for the authors to perform a phylogenetically controlled statistical analysis, but it is important to acknowledge this weakness of the current analysis.  I refer the authors to the immense literature on phylogenetic comparative analyses, which they likely are already familiar with (eg Harvey and Pagel 1991 The Comparative Method in Evolutionary Biology, Oxford University Press).
Validity of the findings: I think the findings are valid, especially if the comparative analyses are discussed in the discussion (see above), and a caveat is added to acknowledge limitations due to 1. inferred pain values and 2. a lack of phylogenetic control.
Additional comments: Minor comments
L19: inert a “While…” or an “…and…” or something else to make this read better.

L115: maybe clarify this a bit.  The current wording made me think that you only have stings available, yet you somehow obtain mesosomal lengths…thus, sting slides are not the only available individuals.  My guess is you did this to avoid destroying samples to get the sting measurements, but maybe make this clearer.

Table 1: i think it would be better if you say in the caption that pain data were not available for all taxa as well, and perhaps have another column indicating whether the pain score came directly from the literature or from personal experience, or inferred from related species.

L229: “ants” should be possessive, either ant’s or ants’, or singular (ant).


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: INFLUENCE AND RELIABILITY OF LOWER-LIMB ARTERIAL OCCLUSION PRESSURE AT DIFFERENT BODY POSITIONS
Review round: 1
Reviewer: 1

Output:
Basic reporting: See general comments
Experimental design: See general comments
Validity of the findings: See general comments
Additional comments: The authors present data on a topic that is of great methodological importance.  This study is straight forward and will be of interest to those implementing blood flow restriction in research and/or clinical settings.  I do, however, have comments that I think will improve the manuscript. My major criticism is there “prediction” equations. I recommend removing them from the manuscript as the important data lies in the change in position. I believe the regression model assumes that the data is normally distributed, which they state it is not.  More importantly, however, is that I do not think the model is stable with such a small sample size. General rules of thumb suggest around 40 people per predictor.  You have multiple predictors but only have 50 individuals. Specific comments below:

Line 51: systolic blood pressure is only a meaningful predictor in the upper body, but the size of the limb is always explaining more variance than any other predictor. This should be corrected.

Line 85: what was the size of the blood pressure cuff?

Line 88: extra period at the end of this sentences

Line 89: it seems more appropriate to just say “skinfold thickness” given that’s what you’re actually measuring.

Line 111: what is the confidence interval around that 0.08?

Line 154: might be useful to add in the variability of the differences into the results

Line 190-203: I feel like this is pretty redundant and could probably be cut down quite a bit

Line 213-214: having references after each one of those suggests that those references demonstrates ischemic reperfusion injury, nerve injury, etc.  Perhaps just state that those things have been speculated in those papers?

Line 217: I think the Jessee and Mattocks paper measured discomfort…not pain per se

Line 220: Martin Hernandez reference would seemingly suggest that volume is only important to a point…not sure that reference really corroborates that sentence completely…consider revising

Line 224: what is a light load?

Line 229: should this say “in a seated…”

Line 231: could someone argue that there is only minimal pressure needed to restrict venous return? I see an argument for manipulating arterial in flow and how that could impact metabolites…but I think this should be made a bit more clear here

Line 225: might be useful to discuss the Ingram et al. investigation…it seems as though that is related to reliability of AOP…and along those lines…how do you separate measurement error from biological variability of blood pressure?
https://www.ncbi.nlm.nih.gov/pubmed/28131507

Line 277: I would remove this entire section

Line 303: how do we know this measurement is “accurate” based on this current study?

Line 308: how much does this measurement actually change? Maybe you only need to measure it once and use that pressure for all subsequent visits?  Would it have to change a decent amount before it meaningfully impacted applied pressure?

Line 317: is there a need to include the company name in the conclusion? Here and throughout the manuscript (outside of the methods)…it might be better to describe it without promoting a product.


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: INFLUENCE AND RELIABILITY OF LOWER-LIMB ARTERIAL OCCLUSION PRESSURE AT DIFFERENT BODY POSITIONS
Review round: 1
Reviewer: 2

Output:
Basic reporting: The paper is well written and easy to follow.
Experimental design: If the cuff was placed at the most proximal part of the leg, why was leg circumference and fat thickness measured at the midway point of the leg? Was this measure located under the cuff for most individuals?
Validity of the findings: I applaud the authors for repeating the 3 measures on a second visit, but I question why the same order that was performed in visit one was not replicated in visit 2. If there is indeed an order effect, then you have added additional error above biological variability/measurement error that can simply be attributed to changing the order.
I applaud the authors for repeating the 3 measures on a second visit, but I question why the same order that was performed in visit one was not replicated in visit 2. If there is indeed an order effect, then you have added additional error above biological variability/measurement error that can simply be attributed to changing the order.

To generate prediction equations, what variables were entered into the model in the first place? You mention that only those that were significant predictors were included in the prediction equation, but which were initially entered? This information should be added.

In the reliability section the only comparison that should be made here is the AOP measured with the Delfi cuff and the AOP measured with the dopplar ultrasound. A comparison of AOP with the Delfi cuff to FMD or MBV measured using ultrasound is not appropriate as these are much more difficult measures and thus will have greater error. If the Delfi cuff could measure FMD or MBV this would be a fair comparison, but only studies using techniques to measure AOP should be compared here. After all, it would presumably be much easier to repeatedly detect the cessation of blood flow than to repeatedly quantify the velocity of blood flow.

Discussion
Line 223: “However, current literature suggests that light-load BFRE training protocols benefit from higher occlusion pressures (80% vs 40%) (Lixandrão et al., 2015), which would support the importance of accurate AOP measurement for prescription of relative pressures.” Another study found no differences between 40% and 90% but this study used 30% 1RM instead of 20% 1RM. Regardless, this study should also be mentioned if this previous study is mentioned
https://www.ncbi.nlm.nih.gov/m/pubmed/26137897/.
Additional comments: The authors compare the necessary occlusive pressure to cease blood flow in three different body positions. Overall, the manuscript is well written and straightforward. Comments are attached.


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: FROM WATER STRIDERS TO WATER BUGS: THE MOLECULAR DIVERSITY OF AQUATIC HETEROPTERA (GERROMORPHA, NEPOMORPHA) OF GERMANY BASED ON DNA BARCODES
Review round: 1
Reviewer: 1

Output:
Basic reporting: Clear English is used throughout, except for some minor grammatical mistakes. I have made suggestions to improve the flow of the introduction.

Literature references and sufficient field background/context is provided.

The article structure and figures look professional. The table could use some work. The Raw data is shared as a text file, but the text (lines 191-196) is missing the Dataset ID and GenBank accession numbers. Custom checks for this manuscript require me to check DNA data availability but I cannot access the deposited data online and the deposition information is not in the manuscript.

Detailed comments:

First paragraph: I would rewrite/reorder this paragraph to make it more apparent why we should care about Heteroptera and give more background on the group. 1) Aquatic insects are the dominant invertebrate taxa. 2) Heteroptera make up __ proportion of aquatic insects and are unique as a group because they contain both aquatic and terrestrial species (compared to mayflies, etc). 3) Different groups of Heteroptera you focus on and their broad distribution. 4) Palearctic region and area of focus for the paper.

The second paragraph needs work. It starts off with a topic sentence about morphological adaptations but covers much more including loss of wings, having wings and migrating, being important components of aquatic ecosystems (but as it’s written it sounds like only the winged taxa are important…), what they consume and their role as prey.

Line 62 – This topic sentence should better fit the second paragraph, perhaps add something about their ecology instead of just having it be about morphological adaptations.
Line 68-70 – different font size?
Line 67 – replace: Furthermore, we observe…, with: We also observe
Line 70 – Replace: Nevertheless, many winged… with: Other winged…
Line 94 – spell out app.
Line 99 – As a consequence, not, As consequence
Line 136 – why are you reporting a result (the number of new barcodes) in the methods section?
Line 136/137 – what are you comparing by using the published barcodes from a previous study? The aim of your study (line 118) should be modified to include all of the goals of your study, including this comparison. Or please explain more clearly how you are using the barcodes from the previous study.
Line 138-140 – Were the specimens from other countries samples that you collected (or got from others) and analyzed for this study, or were these just barcodes that already had been done?
Line 239 – Figure S2 is very hard to read because it spans so many pages, but I’m not sure what the solution is if you include every single individual.
Experimental design: The research is of high quality and the question is relevant and meaningful. Many aquatic insect taxa are difficult or impossible to identify to species in their larval stages (and even as female adults, as described by the authors). The use of barcoding and other techniques to identify species will allow important ecological questions to be addressed that cannot with taxonomic methods alone. Also, the authors even found species in Germany that had not been described before. Perhaps this result should be highlighted a bit more?

The research question is well defined, relevant and meaningful. The authors state how their research fills an identified knowledge gap in the identification of Heteroptera.

The DNA barcoding and analysis sections had good detail. I had a few questions regarding the species collection and identification section (listed above). The methods are described in sufficient detail to replicate.
Validity of the findings: The data are robust. I do wonder if results would be different if more range wide samples of different species were included. In some cases the samples sizes were quite small (i.e. one individual).

The conclusions are well stated and new DNA barcode and relationships between different species are presented. A thorough discussion of species with high intraspecific and low interspecific differentiation is also provided.
Additional comments: This paper provides a nice dataset on the molecular diversity of aquatic Heteroptera. It would be interesting to provide some information or discussion on the additional information that could be gained in a typical study of Heteroptera in Germany by using the DNA barcodes instead of just using traditional taxonomy. Are a large proportion of the individuals captured impossible to identify because they are immature or females. Are certain times of year more difficult to study because of the life stage of the organisms? Perhaps a little more discussion of how DNA barcodes might change our conclusions in ecological and conservation studies? This would be useful additional information in the introduction. Improving the introduction in general (and using the comments from the Basic reporting section) would make this a stronger paper.


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

Input:
Title: FROM WATER STRIDERS TO WATER BUGS: THE MOLECULAR DIVERSITY OF AQUATIC HETEROPTERA (GERROMORPHA, NEPOMORPHA) OF GERMANY BASED ON DNA BARCODES
Review round: 1
Reviewer: 2

Output:
Basic reporting: In this study Havemann et al. work to create a DNA barcode library for 67 species of Heteroptera or true bugs found in Germany. Overall, the manuscript was well written and very clear, though I did find a fair number of small typos, many of the detailed in my general notes. The figures were also very nicely done.

I thought the analysis and results were interesting and likely helpful for other researchers in the field, but I felt the greatest weakness was there was not a clear description of previous studies and findings. The work was also purely a description based study using DNA barcode information. Many of the most intriguing findings were suggested for future work including hypothesis driven questions. Without some description of how these results extend or improve upon previous work, it is hard to assess if this manuscript is answering a specific knowledge gap. I think it could, but it would greatly aid if the authors provided more detail on this gap is and how they are helping to fill it. There are hints of it, but nothing that seems very substantial or convincing, likely because of a lack of detail of how these new information fit into the context of what was known before.

1.  Ln 112-115 It was odd to me that the authors state there have been "few" studies, but then they are able to list 11 other articles including 2 that focused on aquatic true bugs (Castanhole et al., 2013; Ebong et al., 2016). Perhaps the main weakness of the introduction is that it wasn't clear to me how they are building on any of this previous work to help bring new understanding and filling important knowledge gaps.

2. Ln 406-409 You use the term "exclusively morphologically-based taxonomy" within "some" species. But this is very confusing because there is no description of what has been done before? Even though there were 11 other studies that have "analyzed the usefulness of DNA barcodes to discriminate species of the Heteroptera" (lns 112-113).
Experimental design: Overall, there were a few cases where very few samples existed, this seems fine for the goal of this manuscript, however I did have one major concern to do with the species where the vouchers were lost (Ln 397). I was not completely convinced that these results should be included given there were only 2 samples to begin with and now no way of further identification or comparison.
Validity of the findings: 1. In multiple places (e.g., lns, 317-319, 327-328, 338-339, 353-355) there was a   statement similar to the following: "Similar to other previously discussed species pairs with low interspecific distances, more specimens have to be analyzed to validate the species status within this subgenus more in detail."

	My main concern with this statement and the fact it is repeated multiple times is if this work is publishable on its own as there seems to be a lot of missing analysis and areas where additional samples are needed to draw a more clear conclusion or answer questions that are hypothesis based and not a purely descriptive study.

	2. Lns 403-404 "In this study we lay the foundations for a comprehensive DNA barcode library for the aquatic Heteroptera in Central Europe and adjacent regions." This point seems debatable given my comment above. It seems a stretch to say this is a comprehensive DNA barcode library when you are also suggesting a lot of additional work needs to be done to understand fully the relationship between multiple of your species pairs? Can you justify this statement or perhaps remove/change it?
Additional comments: Ln 38-40 This sentence is confusing as worded and also doesn't appear to grammatically correct. Can you rearrange to help clarify? And/or split these 2 ideas up and add a bit more detail for each?

Ln 80 remove one of the "e.g."

Ln 81 "the determination by", I first assumed you meant phenological placement? But the next sentence made me realize you just mean species determination. It might help to clarify this sentence to say exactly what you mean.

Ln 92 "During the last years" doesn't sound correct. Phrased this way, it seems to say the end of something rather than the last few years.

Ln 94 "app." Is this an abbreviation? This should be spelled out, no reason to have an abbreviation here.

Ln 99-100 should be… As "a" consequence and later in the sentence by "a" unique

Ln 155 - There seems to be an inconsistent use of parentheses when using e.g. as other times they were previously not included, but are included in this sentence.

L209 - missing a "the" I think before "model"

Ln 220 - I think the "of" needs to be removed

Ln 222 - need a "an" after "and"

Ln 225 Change to "In the case"

Ln 298 - I think it should be clusters and not "cluster"?

Ln 333-335 "While this character is fairly good for most typical specimens"… fairly good for what? Identification? Also, the word character is used twice in the sentence, but I think you mean characteristic.

Ln 335-336 Not sure the purpose of this sentence, seems a bit anecdotal. Perhaps add more detail relevant to your discussion or remove it?

Ln 338 "only one (female) specimen… analyzed so far", I would suggest perhaps changing this sentence as it sounds like you have additional samples that need to be analyzed. If so, why are they not included in the current manuscript?

Ln 381 "is not scope" should be "is not within the scope"

Ln 403 "for 55 species determined species"? Not sure how this is supposed to read?


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: FROM WATER STRIDERS TO WATER BUGS: THE MOLECULAR DIVERSITY OF AQUATIC HETEROPTERA (GERROMORPHA, NEPOMORPHA) OF GERMANY BASED ON DNA BARCODES
Review round: 1
Reviewer: 3

Output:
Basic reporting: The written text is clear: the story flows, the context and aims are clearly introduced and indicated in the first section, material/methods and results are well structured and unambiguous, and the discussion of the results is not over-extrapolated and opens the door to future research. The literature provided is also context-related. The figures and associated legends are understandable without going back to the main text.
Experimental design: The research question is relevant and well explained: they aim to fill the barcode reference library gap for the two target groups, with a lot of the species’ being bioindicators of water quality. Filling these gaps opens the door to future eDNA studies, like rapid biodiversity inventories, temporal/spatial biodiversity investigations etc. The sampling is large in number and covers the country’s geographical range. A large proportion of the species of the two target groups presently recorded in Germany are represented, most of the time by more than one sample. The methodology is nicely described.
Validity of the findings: Conclusions are well stated, linked to original research question and limited to supporting results. The study provides reference material for future eDNA studies and underlines some limits of the DNA barcoding technic for some closely related species. Data and analyses are robust.
Additional comments: Minor comments:
- line 130: were all the samples collected at adult satge? Did the authors only used the male specimens for DNA barcoding when species identification only relies on male characteristics?
-line 135-136: the sentence should go in the result section
-line 144: not recorded but I assume that they were collected in Germany. If so, please specify in the sentence.
Same line: for how many species exactly did you only had one barcode? were these samples included in the further analyses?
-line 268: h8, could you specify where is this sampling coming from? Was it collected out of Germany?
In this paragraph it might be nice to give some indications about the geographical coverage of each species sampling because the larger it is, the higher the chances to have more haplotypes recorded.
-overall, check for brackets after species name. Sometimes they are present, sometimes not.


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: FROM WATER STRIDERS TO WATER BUGS: THE MOLECULAR DIVERSITY OF AQUATIC HETEROPTERA (GERROMORPHA, NEPOMORPHA) OF GERMANY BASED ON DNA BARCODES
Review round: 2
Reviewer: 1

Output:
Basic reporting: I greatly appreciate the authors revision in taking into consideration my major comments and addressing each of them both in the introduction and in the discussion, as well as answering my questions. I feel the authors have done a thorough job in motivating the need for their study, including the major knowledge gaps and how the results of their current study will spur future research. I found a number of small typos that I detail under general comments. Well done, overall.
Experimental design: No comment.
Validity of the findings: No comment.
Additional comments: Ln 107: "Since a few years", suggest something like, "In the last few years,"

Ln 107: change "approaches become" to "approaches have become"

Ln 108: I think you mean something like "overcome possible drawbacks of traditional ways of assessment". What do you mean by the traditional way of assessment? Taxanomic?

Ln 119: Switch from "highly relies" to "relies highly"

Ln 123: Remove the extra period

Ln 126: Needs an "a" in front of "unique"

Ln 128: Needs a "the" in front of "global"

Ln 155: There is an extra period after "Cameroon"

Ln 393: "expect" should be "except" I think

Ln 475: change from "got lost" to "were lost"

Ln 477: there is a typo in "tothe"

Ln 483: Looks like a typo, not sure what word is missing here "Unique BINs were revealed for 55 species determined species", also further in the sentence, you can drop the "of" from "representing of"


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: SALT MARSH SEDIMENT BACTERIAL COMMUNITIES MAINTAIN ORIGINAL POPULATION STRUCTURE AFTER TRANSPLANTATION ACROSS A LATITUDINAL GRADIENT
Review round: 1
Reviewer: 1

Output:
Basic reporting: This manuscript is clear, concise, and well-written throughout.  The authors provide the appropriate context of the work and reference the relevant literature.  They background sets the stage for their work and provides a rationale for why the work is important. Their hypotheses are clearly articulated and testable. The methods are reasonable and their data analysis seems appropriate for the questions they are asking.

 Just a couple minor points:

It might be nice to include in your methods or results, some description about the plant side of the reciprocal transplant.  Although you do mention it in the discussion, I did find myself wondering as I was reading your results whether the plants survived the transplant or if there were inhibited from being successful in their new system. It seems from the results that the northern marshes did poorly in the southern system but the southern marshes did fine up north?  It might be worth bringing that information in earlier and adding a plant-soil feedback context.

Line 407 - liner should be linear.
Fig. 1 legend - first, figure 1 is actually not referenced in the text. Second, the legend is written essentially as a statement of the main conclusion of the findings, though I assume that the map will be introduced earlier  - I looked at the legend and assumed that the figure was going to indicate the effect of transplantation when really it is only a map of sites.  Perhaps make the legend a bit more congruent with the content of the figure.

Fig. 3 - I found the P really hard to see in some of these - could the authors use colors instead, to differentiate?  It also might be useful to have the northern marshes be open symbols and the southern marshes be closed symbols to make that contrast more clear.
Experimental design: The experimental design for this experiment was straightforward, well described, and clear.  All data analysis seems appropriate.  I liked the matrix approach.

The authors took sediment cores from five marshes and did a full reciprocal transplant, with all proper controls. They left the transplants out for a growing season and then harvested them to see if environmental factors or dispersal play a dominant role in structuring microbial communities.
Validity of the findings: The results are interesting.  Despite a huge latitudinal gradient, the microbial community of transplanted plants retained the signal of there native lands after six months. I wish there were three more years of data to see how long this pattern persisted!  I do think a little bit of context for how the plants did would be useful for understanding the broader picture.

Conclusions are not overstated and are well supported by the data.
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: SALT MARSH SEDIMENT BACTERIAL COMMUNITIES MAINTAIN ORIGINAL POPULATION STRUCTURE AFTER TRANSPLANTATION ACROSS A LATITUDINAL GRADIENT
Review round: 1
Reviewer: 2

Output:
Basic reporting: The article is well-written, clear, and fully cited throughout. The figures look fine, though some larger text could help in some legends. The MS is self-contained as well.
Experimental design: The experimental design meets PeeJ's scopes/aims. The questions are well defined, relevant, and meaningful. The research does address a knowledge gap in salt marsh microbial ecology.
Validity of the findings: The findings are valid, though I think more discussion of the nuances of eh experimental setup is needed. The stats and data are robust and the experiment is well replicated.

I think there's a bit too much speculation in the discussion and could be grounded more in the ltierature.
Additional comments: The study by Angermeyer and colleagues surveys the response of salt marsh sediment bacterial
communities to a large reciprocal transplant experiment between several marshes. They find that the
communities are resistant to transplantation which may suggest limited dispersal of the communities.
This study is well-written, interesting, and the results are generally clear but do need some clarification
at spots. Below I list some major comments and then below I detail some more specific comments
relevant to the text.

Major comment: what ecotype of Spartina was selected for this study? Was it short or tall? This
ultimately has important implications for their study and the interpretation of the data due to the
starkly different habitats that the to ecotypes reside in. If this is the tall ecotype of Spartina then the
plants are experiencing tidal flooding every day, whereas the short ecotype only sees flooding 1/3 of the
month (e.g. Johnson et al. 2016, Ecol Appl).
Given this, if tall was selected the authors are likely seeing full integration of the transplanted soil into
the local conditions of the new marsh. If short was selected, then I do not believe the transplanted soil
fully experienced the abiotic conditions of the marsh itself (e.g. Balser and Firestone 2005, Biogeochem)
and the small change in temperature between sites (17-24C) likely not enough of a push to drive
changes. If the short ecotype was selected the authors would need to demonstrate the exposure of the
transplanted sediments to the new conditions with some measurements of the soil abiotic conditions to
convincingly demonstrate the efficacy of the transplant.
Related to this, I wonder how much exposure to new conditions sediment 20cm deep in marsh sediment
receives. Unless these are bottom up flooding marshes (like many in New England are), these sediments
likely wouldn’t see that drastic of change in their abiotic conditions in particular because the biggest
deciding factor here (air temperature) likely isn’t all that differ 20cm deep.

Title: temperature is just one component of what makes up the gradient sampled here, especially in
terms of the soil chemistry. I would suggest revising the title to reflect this. Also, population is the wrong
word here, community is more appropriate. Something like “Salt marsh bacteria communities maintain
structure despite transplantation’.

Methods
General comment: the authors should probably remove Chloroplasts from this analysis. V4 is generally
terrible for this group of taxa and the short-read length makes meaningful identification of these
eukaryotes difficult at best. Also, why mantel tests in lieu of something like a
PERMANOVA/PERMDISP/ANOSIM?
Line 139: did this include a plant(s) or was it just bare sediment?
Line 140: it would be helpful to know a bit more about where in the marsh they were transplanted.
Things like height relative mean sea level, tidal inundation, distance from the creek bank, and a bit more
about the marshes themselves to better interpret the experimental design.
Line 151: I assume this is just something like cheese cloth, but it would be helpful to know the mesh size
here.Line 155: So the authors bored 20cm into the sediment with a spatula? How are they sure they reached
the desired depth and didn’t get mixing of higher layers? The sampling says 20cm, but was it around
20cm (e.g. 19-21cm)?
Line 178: why the DNA/RNA kit when only DNA is presented?
Line 188: delete ‘Benchtop’.
Line 190: I would find it helpful for the authors to just include what they did instead of having to go to
GitHub/another paper.
Line 192: data was rarefied before OTU picking? This seems odd and potentially biasing OTU clustering
by removal of diversity. Can the authors please clarify?
Line 193: OTU picking method? Closed pick? De novo pick?
Line 222: this is on rarefied data, no? Absolute abundances in this instance seem to be perfectly viable
over relativized abundance.
Line 237: It would also seem more appropriate to use a ANCOVA for the environmental variables and
OTUs, holding origin and site as effects. Given that the variables change as a function of site and covary,
this should be taken into account in the regression model.
Line 241: Bonferroni corrections are notoriously overly conservative, did the authors try another p-adjustment technique like Benjamini-Hochberg?

Results
Line 250: remove ‘host’
Line 251: I also assume no roots entered the cores as well?
Line 258: Was anoxia measured? Creek bank sediment is only usually anoxic during flooding.
Line 275-276: is this information about DNA mass needed?
Line 283: this paragraph could be strengthened with some statistics (e.g. Kruskal-Wallis test w/pcorrection).
Line 284: Drop the ‘s’ from the end of ‘compositions’.
Line 287: not a requirement, but oligotyping would be interesting for this Vibrio OTU.
Line 314 (and in methods too): is there a reason the ordination would need to be done with all OTUs and
then again with the top ten most abundant taxa? Bray-Curtis is most strongly influenced by the top taxa
and given that the ordinations look very similar, I question whether it is needed because the authors do
not do anything with these top ten taxa.
Line 321: indicator OTUs is misleading here. This implies that they are indicator species, as determined
by that specific test. I don’t disagree with how the authors determined these four OTUs, however I’m
surprised that only 4 of the 21k OTUs were significantly correlated.

Discussion
Line 336: this paragraph rehashes the introduction.
Line 345: one thing that’s hard to tell about this experiment is whether dispersal, due to tidal flooding,
occurred especially at the depth sampled. Do the authors have data about the sediment chemistry
outside
Line 346: this is not just a temperature gradient; many things change along the eastern US coast.
Line 350-352: without presenting more data about the sites (e.g. porewater chemistry, ocean water
chemistry, variations in abiotic conditions, ect), this is a hard claim to make. The physical properties of
the sediments themselves, as noted in the methods/results, suggest these sites are very different.
Line 355: this implies that sampling didn’t disrupt the communities, not that the methods were effective
at excambing the question. Which ultimately is a strength given the likely release of carbon via severing
marsh roots.
Line 362: while this intuitively makes sense, the authors lack the resolution in terms of measured
variables to really parse out the driving factors here.
Line 365: remove ‘host’.
Lines 368-370: this sentence is confusing, I’d rephrase or remove it because the next sentence says all
that’s needed.
Line 378: one thing about this paragraph that the authors do not discuss is the idea of no mixing of the
local conditions. This has been discussed by several papers (e.g. Balser and Firestone 2005, Soil Biochem;
Hughes and Martiny 2013, ISME J; Gasol et al. 2002, Limnol Oceano). Given this, it is likely that their
results show no mixing. In addition, a recent paper about dispersal in soil systems would also be
appropriate for discussion (Evans et al 2017, ISME J).
Line 402: while Vibrio do bloom in the summer, the authors sampled in October. Work by Bowen et al
2009, ISMEJ showed seasonal patterns in the microbial communities in marshes, which was also show in
the active communities by Kearns et al. 2016, Nat Commun. Given this, how confident can we be that at
the end of the season samplings are reflective of the warmer temperatures?
Further, given the lack of change in transplanted cores it suggests its likely something about the soil
chemistry instead of the temperature. This may be reflective of the sandy nature of southern marshes
relative to the high carbon marshes in the north.
Line 438: the presence of Vibrio regardless of temperature suggests lack of mixing, which may be why
the persist. I also think that the idea of resistance, which has been shown a few times for marsh
communities (Bowen et al. 2009, 2011, ISME; Kearns et al. 2016, Nat Commun; Angell et al. 2018, Front
Micro), is an interesting talking point here.
Line 458: while I find this paragraph interesting, I question how applicable it is to this study given that
the majority of studies in this area of plant-microbe interactions, outside of the work of Bowen and
colleagues, shows the environment overrides plant genetic effects every time. Further, based on the sampling, it appears this study would be independent of the plant itself since the authors do not
mention roots and the rhizosphere effect only extending a few millimeters from the root surface.
One other point about this is that Phragmites is hardly a salt marsh plant as its home range is in brackish
(<10ppt) to freshwater environments, which all Spartina would be unable to compete in.

Figures
Fig 1: looks good. Perhaps scale bar?
Fig 2: I would recommend removing the failed samples, its not needed and it distracts from the figure.
Also the size of the text could be improved.
Fig 3: the shape/letter makes these hard to interpret at first site. Perhaps color them?
Fig 4: I would add a legend, it’s a pint to look back and forth between this figure and figure 2.
Table 1: are these averages over the course of the experiment or just a one time point measurement? If
an average, please include the SE or SD.


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: IS THE FUTURE ALREADY HERE? THE IMPACT OF CLIMATE CHANGE ON THE DISTRIBUTION OF THE EASTERN CORAL SNAKE (MICRURUS FULVIUS)
Review round: 1
Reviewer: 1

Output:
Basic reporting: No comment
Experimental design: From line 216 to 224
It is better if you could give the values for the following MaxEnt parameters which you have used in your model
1) Regularization multiplier
2) Maximum iterations
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: IS THE FUTURE ALREADY HERE? THE IMPACT OF CLIMATE CHANGE ON THE DISTRIBUTION OF THE EASTERN CORAL SNAKE (MICRURUS FULVIUS)
Review round: 1
Reviewer: 2

Output:
Basic reporting: The manuscript utilizes a popular method to study climate driven range shift pattern of snakes in the US. This is an important contribution considering not much work has been done on these specific reptiles using this method in the past. The manuscript is written well and included methodology which utilizes established methods for assessing model accuracy for present and future conditions.

My specific comments on the sections listed below in sections with lines numbers

Line 105: Language could be improved to ensure the audience can understand the text, for example in this line, do the authors mean “evaluating the utility of future climate models in relation to recent coral snake occurrence prediction in the years between 2001-2015”?
Experimental design: Line 108-109 & 216-224: Model environment settings were not mentioned (i.e. Authors did not mention whether they used default settings of MaxEnt or modified some of the parameters and whether it was executed trough the java applet or through R).

Line 149: Although it was explained the utility of the buffer but nothing mentioned about why 200 km buffer was generated not 100 km or 50km? More details about this would be helpful for the readers.

Line 221: In this type of modeling studies it is expected the model repeated at least 100 times for ensuring the robustness of the result. Also bootstrapping in Maxent with observation data split into training and test run is often used for the replicate runs to add additional layer of validation trough AUC’s. Authors need provide justification why model was only repeated 10 times and without any data split.
Validity of the findings: no comment
Additional comments: Additional figures could be added in the appendix showing all the environmental layers used for the modeling which will help readers to understand whether the distribution could be derived by particular environmental variable or with combination of them.

I thank the authors for providing the raw data but including a descriptive metadata would be useful for the readers and also raw data of ‘locality points 2000-2015’ file need to have ‘Species’,’ data source’ fields like other location file.

Authors could consider the following citations to enhance their introduction and discussion section
Ceia‐Hasse, Ana, et al. "Integrating ecophysiological models into species distribution projections of European reptile range shifts in response to climate change." Ecography 37.7 (2014): 679-688.

Araújo, Miguel B., Wilfried Thuiller, and Richard G. Pearson. "Climate warming and the decline of amphibians and reptiles in Europe." Journal of biogeography 33.10 (2006): 1712-1728.

Chen, I-Ching, et al. "Rapid range shifts of species associated with high levels of climate warming." Science 333.6045 (2011): 1024-1026.


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: IS THE FUTURE ALREADY HERE? THE IMPACT OF CLIMATE CHANGE ON THE DISTRIBUTION OF THE EASTERN CORAL SNAKE (MICRURUS FULVIUS)
Review round: 1
Reviewer: 3

Output:
Basic reporting: This paper studied the present and future distributions in the eastern coral snake (Micrurus fulvius) in the United States. They also compared the current and future distribution models to see if the distribution range shifts are already happening in this specie or not.

This paper has some important results to science where there is a gap of knowledge on climate change impacts on future distributions on reptiles. However, I believe that this paper in its current format is not publishable and needs careful and considerable revisions. Please find my comments in details as follow and also in the attached annotated PDF file.
Experimental design: Introduction
The structure of the paragraphs are not following each other. Most of the paragraphs do not have a main topic and are just some general introduction.
Although you mentioned three objectives for your research, but still the importance of doing this research and the benefit of the outcomes are not clear enough. You need to mention what is the main gap of knowledge and how your study addressed this. What would be the benefit of your future models to establish management plans for the areas of future inductions and so on.
You also need to talk about the physiology, ecology, and habitat preferences of the coral snakes. These information are missing in the discussion as well.

Methods
Some methods which have been used in this study need to be explained more in details in the Methods and also the Methods need to be shortened and be more concise. I mentioned some examples both in the annotated PDF and as follow:

1- you need to explain why you chose these environmental factors. You should explain clearly the role and importance of each layer in biology, physiology, or ecology of the coral snakes.
2- you should add a specific part to your method explaining the most important environmental factors in creating your present and distribution models. This part is very important to explain the models based on the contribution and importance rates of the layers you used.
3- you have to include more information for the data, e.g., Unit, Type, Temporal range, Minimum, Maximum, Mean, Std. Dev.
Validity of the findings: I would like to see the response curves or at least a table which shows the contribution and importance rate of each layer in both present and future models. This part is really missing from the paper and it is one of the most important outcomes of the Maxent modeling.
Additional comments: 1- please use more recently studies on climate change modelling using Maxent and compare your result with those ones. There are many studies have been done since 2014 which many of them are missing from your reference list.
2- please minimize the use of one-off reference in your general statements. Once you are talking a bout a fact and staying many studied found it, you should at least give 2-3 references, please refer to the annotated PDF.
3- please also divide the discussion in two parts, present and future modeling. In the current format, discussion is not concise and very hard to follow what you mean in many of the paragraphs. At the end of the Discussion, I was left with a "so what" question.
4- please revise the formatting of the reference list, there are many typos, e.g., lines 550 (issue No. in Bold), 668 (year in Bracket), and 729 (issue No. is missing).


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: IS THE FUTURE ALREADY HERE? THE IMPACT OF CLIMATE CHANGE ON THE DISTRIBUTION OF THE EASTERN CORAL SNAKE (MICRURUS FULVIUS)
Review round: 2
Reviewer: 1

Output:
Basic reporting: This manuscript is a thorough study of the impact of climate change on the distribution of the eastern coral snake (Micrurus fulvius). MaxEnt (version 3.3.3. k) species distribution modeling software was used to evaluate range shifts from 1950 to 2000 and 2050 of the eastern coral snake. The authors report that the future climatic conditions will increase the habitat suitability across the range. They predict the Southeastern United States will have a novel environmental condition in 2050. This paper has important findings for reptile conservation and in biogeographical studies.
Experimental design: Thank you for making changes in line 220-222.
The study has been carefully designed and the steps of the methods are clear and easy to follow. Nevertheless,  I have two additional general comments:
1)	Line 160 & 277: The conversion of arcminutes to km. The given conversion is incorrect. It must be approximately 4.6 km at the equator.
60 arcminutes = 1 degree = approximately 111 km at the equator.
5 arcminutes = 9.2 km at the equator.
2)	Line 220 -222.
There are many successful studies which have used cross-validation and 10 replicates. Please add references and provide a basic description of cross-validation.
Validity of the findings: Thank you for providing supplementary files.
It is difficult to see whether the resulting MaxEnt model maps have a higher prediction for the true natural habitats (true presence) from 1950 to 2000.
I would like to suggest to change the color of the maps in figure 2. It is better if you can reclassify these resulting maps using a mapping software and give contrast colors for each class. After that add occurrence records to the same map for the better visualization.
Additional comments: No comment