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: ACUTE ISCHEMIC PRECONDITIONING DOES NOT INFLUENCE HIGH-INTENSITY INTERMITTENT EXERCISE PERFORMANCE
Review round: 1
Reviewer: 3

Output:
Basic reporting: The purpose of this study is to exam the acute effect of ischemic preconditioning (IPC) on a high-intensity intermittent exercise performance and physiological indicators in amateur soccer players.  Paper is poorly written and lack of clarity in some sentences (please see below for the details).  In addition, there are a lack of details in methodology (for example, what is the sampling time for lactate?). Lastly, there is a lack of depth in discussion.
Experimental design: According to the previous studies (Patterson et al. 2015 & Incognito et al. 2016), IPC does demonstrate that there is an improved peak and mean power output during the early stage of repeated sprint event as well increase the maximal oxygen consumption during incremental bicycle exercise. My understanding is that the time interval between IPC and exercise testing should be essential in the impact of the IPC. However, there is a lack of information of the time interval between IPC and Yo-Yo intermittent endurance test.

Although this study included three groups (IPC, SHAM, Control) with single-blinded design, it is still very different to rule out the psychological effect of the participant. The significant finding form the previous study could be partially explained by the highly motivated participant following IPC.

Since there was IPC non-responder and responder individual difference in soccer players, having a larger sample size might make more sense.

Incorporating EKG, ventilation, R , VO2, peak power,  and maximal oxygen measurements  in this study will strengthen the quality of this paper.
Validity of the findings: no comment
Additional comments: Introduction
Line 51-59: there is a lack of clarity in these sentences. For example “ being associated with both VO2 and citrate synthase activity in untrained men” I am not sure what this means?  Based on the logic flow, author assumed that untrained is similar to amateur which I am not completely agree with.

Discussion:
There is a lack of discussion in depth in physiological explanation for the absence of significant finding during YoYoIE2 test following IPC in comparison to other studies, although author briefly mentioned that this difference is probably due to the type of exercise. It seems like that IPC had a certain acute effect in exercise performance from previous study, however how long this acute effect could last was not clear. The nature of the intermittent maximal test might counterbalance the acute effect following IPC.


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: ACUTE ISCHEMIC PRECONDITIONING DOES NOT INFLUENCE HIGH-INTENSITY INTERMITTENT EXERCISE PERFORMANCE
Review round: 2
Reviewer: 1

Output:
Basic reporting: The authors have addressed my major concerns.
Experimental design: See below
Validity of the findings: See below
Additional comments: The authors have addressed my major concerns. I accept it in the latest 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: ACUTE ISCHEMIC PRECONDITIONING DOES NOT INFLUENCE HIGH-INTENSITY INTERMITTENT EXERCISE PERFORMANCE
Review round: 2
Reviewer: 2

Output:
Basic reporting: The study investigated the acute effects of ischemia preconditioning on high intensity intermittent exercise performance in amateur soccer players. The authors found out that IPS had no influence on the exercise performance.
Experimental design: The ischemic preconditioning protocol is under the steady state where muscle was not working and in the rest condition. Please consider to include some level of exercise in the ischemic preconditioning protocol that may generate better preconditioning effects.
Validity of the findings: OK
Additional comments: NO


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

Input:
Title: ABILITY TO PREDICT REPETITIONS TO MOMENTARY FAILURE IS NOT PERFECTLY ACCURATE, THOUGH IMPROVES WITH RESISTANCE TRAINING EXPERIENCE
Review round: 1
Reviewer: 1

Output:
Basic reporting: Please see below
Experimental design: Please see below
Validity of the findings: I believe the statistics need to be re-run.
Additional comments: The authors have investigated an interesting idea, however, I believe the authors may need to re-calculate all of their values. For example, it’s hard to believe that a mean difference of 4 repetitions corresponds to a standard error of 1.  I downloaded the spreadsheet and used your data and I believe that you put the “predicted” in the criterion instead of putting the “actual reps” as the criterion.  Please see below.  Also, you may consider implementing the minimal difference (SD of difference score x 1.96) in addition to what you have. I believe that these changes will likely change some of the interpretation of the manuscript.

Just a couple of examples,
Beginner Chest Press
You have 1.94 (1.48, 2.84)
I believe it should be 4.0 (3.04, 5.84)

Leg Press Expert
You have 1.27 (1.05, 1.63)
I believe it should be 1.73 (1.42, 2.21)

I would also delete the figure, it adds nothing to the manuscript.

A few other things to consider:
Introduction: Resistance training is associated, at best, with a wide range of health benefits…this should be made quite clear.  The Dankel study does not actually offer much support for “resistance training” as it found the strongest associations with the outcome of strength as opposed to the behavior.  I would tone this down some

Was this IRB approved? This information should be put into the manuscript.


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

Input:
Title: ABILITY TO PREDICT REPETITIONS TO MOMENTARY FAILURE IS NOT PERFECTLY ACCURATE, THOUGH IMPROVES WITH RESISTANCE TRAINING EXPERIENCE
Review round: 1
Reviewer: 2

Output:
Basic reporting: Quite a negative tone concerning the accuracy and use of repetitions in reserve (RIR) during resistance training. Therefore, the writing style does not seem to present a balanced argument of RIR for resistance training.
Experimental design: The investigation performed is not adequate for the aim of the study. The investigators did not have participants estimate repetitions in reserve (RIR) during the actual exercise used to assess accuracy in RIR.Therefore the results presented are highly questionable.
Validity of the findings: There are a lot of unknowns that affect the results reported in this study, but the major one relates to the flawed methodological design.
Additional comments: Overall this paper is written in quite a negative tone concerning the accuracy of using repetitions in reserve (RIR) for monitoring of resistance training effort. An error of approximately 1 repetition is excellent for training purposes and RIR is best method that can be used to monitor resistance training effort. For example, RPE during resistance training is poorly correlated with RIR (soon to be published paper) and the information derived from RIR is impossible to fully decipher. So RIR is the obvious best method to provide coaches and trainers with added information, but RIR should not be used to prescribe resistance training. The RIR is most useful for assessing progression (adaptations) and fatigue (predictor of overtraining and maladaptation).

The sample size used for this study is great but there is a major methodological flaw. The accuracy of the RIR can only be assessed with a trainer reporting their rating during the exercise in which they will then performed repetitions to momentary failure. This is because you need the trainer to be able to utilise the many physiological and psychological cues to make their estimation. It makes no sense to have trainers make an estimation based on their previous training experience. The number of repetitions performed to failure with a specific relative load is likely to change on a day to day basis. Factors such as sleep, diet, previous training, time of day, other stressors, motivation etc. will impact upon the repeatability of performing repetitions to failure at a set load. The participants in your study may not have been motivated by the assessor but they were under research conditions and probably confounded your results. Also, it is no surprise that you found more experienced trainers would be more accurate with RIR. I might even hypothesise that this was due to the experience RT greater training frequency. Did you assess when your participants performed their last training sessions prior to your study? Potentially this could have impacted some of the results.

Specific concerns:
Line 65: ‘applications of RT to maximise these outcomes …..’ does not make sense?  The ‘application’ refers to putting RT into practice. I think you mean the RT prescription?
Line 106-108: Trained participants are good at predicting repetitions to momentary failure. What criteria are you using to make your statement? You could not be possibly thinking that trainers can develop an ability to predict RIR with 100% accuracy. It will always be dependent on the loads used and proximity to momentary failure.
Line 194: Never end sentence with a prepositional primer (also line 316).
Lines 232-233: Anchoring of perception…… interesting because the study participants were making estimates based on previous exercise experience. Seems inappropriate to have this discussion


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

Input:
Title: ABILITY TO PREDICT REPETITIONS TO MOMENTARY FAILURE IS NOT PERFECTLY ACCURATE, THOUGH IMPROVES WITH RESISTANCE TRAINING EXPERIENCE
Review round: 1
Reviewer: 3

Output:
Basic reporting: No comment, all areas of basic reporting meet standards.
Experimental design: No comment, all areas of experimental design meet standards.
Validity of the findings: Data is robust and statistically sound and controlled. Interpretation is the only area where revision is recommended; see general comments.
Additional comments: First I wish to thank the authors for their contributions to this field of study. This is an important body of evidence to inform this area of research, the writing is clear, the findings are well presented, and the methods are sound. Additionally, the authors are correct that the work by Hackett and colleagues is influenced by the fact that multiple sets to failure were performed in both their 2012 and 2016 studies. Thus, as correctly pointed out by the authors, previous sets would help to improve accuracy of subsequent sets, thus potentially inflating the statistical representations of accuracy in both studies. Thus, the approach of estimating how many repetitions the lifter believes they can accomplish before ever beginning a set, and then comparing that estimation to the actual performance is novel.

That said, there are some revisions needed to ensure an accurate comparison of this data set is made to that of the existing literature.

In the title, and lines 41 (abstract), 107, 197, 202, 235, 261, 264, 276, 278, and the 318, the use of the word “poor” should be amended to a less subjective term. I would advise “not perfectly accurate” as this isn’t a debatable description, given the subjects in this study were unable to predict with complete accuracy how many repetitions they could perform to failure with a given load. The reason I believe this is important is the authors cite Hackett 2012 and 2016, and refer to the accuracy as “poor”, while Hackett and colleagues themselves in 2012 referred to the same accuracy as “high” and then as “quite good” in 2016 in the discussion sections of both papers. Specifically in their 2012 article, they state the following “Despite the differences between the estimated- and actual- repetitions-to-failure for the earlier sets of exercise, the difference was approximately only one repetition, indicating that participants only slightly underestimated this.” The use of “only slightly underestimated” contrasts the authors’ use of the word “poor” to highlight my point.

Both Hackett and colleagues and the present authors use a subjective description for the objective distance to failure achieved by their subjects. Neither can be said to be “correct” as it is a subjective determination, however, the authors can use less subjective terminology. Thus, please change all instances of “poor” that I highlighted above to “not perfectly accurate”, or similar, or when in doubt, simply state the actual distance from failure quantitatively as the number of repetitions.

The limitations the authors refer to when citing Hackett 2016 in lines 111-116 need to be revised as they contradict the methods of Hackett 2016. As it is stated in the methods of Hackett 2016 “Subjects aimed to perform 5 sets per exercise, but if concentric failure occurred prior to reaching 10 repetitions for a set, the testing for the exercise ceased.“ Additionally, in the beginning of the results section of Hackett 2012 it is stated “For both the bench press and squat, the estimated- and actual-repetitions-to-failure for set 5 was 0 for all participants. Therefore, data from set 5 were excluded from the ANOVA.” Based on these two quotes, the authors should revise the stated limitations of the data by Hackett and colleagues as it doesn’t appear that sets were continued if failure was reached at or before repetition 10, and if failure was reached at repetition 10, the estimation of 0 repetitions remaining was not included in the analysis.

Finally, I think the authors do a fantastic job pointing out the issue of an a-priori assumption of accuracy and of the short comings of RIR as a resistance training prescriptive tool. However, on balance, I think the authors should also point out some of the benefits of RIR based load prescription. One, that the authors already point out, is that it is perhaps more appropriate for strength athletes and bodybuilders vs untrained individuals. Indeed, the Hackett 2012 paper showing ~1 repetition away from accuracy was on bodybuilders, and almost the entirety of work by Zourdos and Helms is on competitive powerlifters. I think that should be specifically pointed as both a limitation for general population users, and potentially a benefit for strength and physique athletes who are much less likely to self-select loads that will not be challenging or lead to adaptation.

Also on balance, please point out what the RIR approach improves on. As the authors pointed out in lines 210-213, studies using traditional Borg RPE sometimes report RPE values in the 6-9 out of 10 range (depending on the study) even when training to muscular failure. Thus, as was shown in Hackett 2012, RIR is likely an improvement Borg RPE for resistance training. Additionally, the authors also point out in lines 87-88, there is large variation in the number of repetitions that can be performed at the same relative load (% 1RM) between individuals. In fact, these differences can be quite large; in endurance athletes compared to strength athletes, on average, a difference of 22 reps at 70%, 8 reps at 80% and ~4 reps at 90% of 1RM https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4042664/. Thus, it would seem that even with the inaccuracy found in the present study, RIR is likely to provide a load closer to the intended difficulty than percentage 1RM based load prescription as well.

To conclude, excellent work, this data is an important contribution collected in a novel manner, and I would recommend the following amendments in order of importance as:

1. Changing the subjective wording of poor to “not perfectly accurate” or similar, or just stating distance from failure when referring to the present and previous studies (by Hackett)
2. Amending the limitations stated about the Hackett paper.
3. Including the fact that despite its limitations, RIR is likely a better alternative than %1RM or Borg RPE for load prescription in resistance training.
4. Adding additional detail to the greater utility for trained individuals, specifically strength and physique athletes as the subjects in papers by Helms and Zourdos and Hackett 2012, were powerlifters and bodybuilders respectively.


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: ABILITY TO PREDICT REPETITIONS TO MOMENTARY FAILURE IS NOT PERFECTLY ACCURATE, THOUGH IMPROVES WITH RESISTANCE TRAINING EXPERIENCE
Review round: 2
Reviewer: 1

Output:
Basic reporting: The authors have substantially improved the manuscript…I have only a few more points
1) you use a comma instead of a “.” In the Table for chest press…please be consistent

2) I believe some of the references you deleted in text still show up in your reference list

3) you say repeatedly that people are “not perfectly accurate”…but you never really mention what would be acceptable? What would be considered good..1 rep? It seems unlikely that you’d ever predict it perfectly?
Experimental design: See above
Validity of the findings: see above
Additional comments: see above


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: ABILITY TO PREDICT REPETITIONS TO MOMENTARY FAILURE IS NOT PERFECTLY ACCURATE, THOUGH IMPROVES WITH RESISTANCE TRAINING EXPERIENCE
Review round: 2
Reviewer: 2

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


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

Input:
Title: ABILITY TO PREDICT REPETITIONS TO MOMENTARY FAILURE IS NOT PERFECTLY ACCURATE, THOUGH IMPROVES WITH RESISTANCE TRAINING EXPERIENCE
Review round: 2
Reviewer: 3

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: I wish to thank the authors for considering all of my proposed revisions. I have no further suggested revisions, and I humbly suggest the paper is now improved. Well done!


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

Input:
Title: ABILITY TO PREDICT REPETITIONS TO MOMENTARY FAILURE IS NOT PERFECTLY ACCURATE, THOUGH IMPROVES WITH RESISTANCE TRAINING EXPERIENCE
Review round: 3
Reviewer: 1

Output:
Basic reporting: The authors have adequately addressed my concerns.
Experimental design: The authors have adequately addressed my concerns.
Validity of the findings: The authors have adequately addressed my concerns.
Additional comments: The authors have adequately addressed my concerns.


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

Input:
Title: ORDINARY KRIGING VS INVERSE DISTANCE WEIGHTING: SPATIAL INTERPOLATION OF THE SESSILE COMMUNITY OF MADAGASCAR REEF, GULF OF MEXICO
Review round: 1
Reviewer: 1

Output:
Basic reporting: In my review of the document, I went through and made quite a few modifications of sentence structure and grammar. The paper is nearly there, but the overall quality of the writing would be well-served by another edit by a native English speaker.

The authors should add the complete R code used in their analyses to the supplementary files available for download.

The authors did a fine job structuring the paper and creating some useful, interesting figures. I would like to see a more explicit hypothesis (re: IDW vs. OK) retroactively suggested in the introduction of the paper.
Experimental design: The results are interesting and useful for spatial ecologists considering IDW vs. OK procedures. As the authors stated, these methods are infrequently utilized in marine studies.

As stated above, the authors need to include their code with their other materials so readers can replicate their work.
Validity of the findings: The authors did a good job outlining the impacts and ecological rationale of their findings. I would like to see the Discussion fleshed out with a paragraph added regarding why the IDW outperformed the OK methods.

The authors successfully and frequently related the findings of this study to those of relevant prior studies.
Additional comments: I enjoyed this paper and hope to see it published. I would like to explore your R code out of my own interest in these methods and curiosity about how they perform in another study region.


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: ORDINARY KRIGING VS INVERSE DISTANCE WEIGHTING: SPATIAL INTERPOLATION OF THE SESSILE COMMUNITY OF MADAGASCAR REEF, GULF OF MEXICO
Review round: 1
Reviewer: 2

Output:
Basic reporting: The paper is very well written and strcutured, has great figures and is generally a joy to read. It makes sense and is coherent.
Experimental design: The experimental design works, and I only have one query: there are a lot of sampling points - Madagascar Reef clearly is the best sampled reef in the world! It would be useful to undertake a sensitivity analysis to find out how many points are needed to get this level of predictive capacity - this would be informative for places with fewer survey points, and also highlight how much sampling effort is required with interpolation to get good predictive capacity.
Validity of the findings: no comment, the paper is very well rounded and the findings are valid
Additional comments: Spatial interpolation methods generally are weak or unsuitable to support spatially explicit analysis in ecology and conservation. This is why i was extremely skeptical about this article, but you succeeded in convincing me that this is a great contribution. My initial reaction also points to an area of improvement: the abstract and early introduction focus on interpolation methods, but do not explain clearly enough under which circumstances they might indeed be useful. Much spatially contiguous environmental data comes from satellite data and has a given resolution, and for projects that want to spatialise survey data at smaller scales, interpolation might be useful because environmental data that could serve as predictor variables to create spatial models do not exist.
I would also recommend to discuss a little more under which circumstances interpolation would be preferred over predictive spatial models.


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: ORDINARY KRIGING VS INVERSE DISTANCE WEIGHTING: SPATIAL INTERPOLATION OF THE SESSILE COMMUNITY OF MADAGASCAR REEF, GULF OF MEXICO
Review round: 1
Reviewer: 3

Output:
Basic reporting: I suggest the basic reporting can be improved. It is often ambiguous and intended messages can be unclear. The English can be improved.

The literature references and background context are good. The introduction was clear and well structured for the study. However, I suggest some of the methods is more appropriate to the discussion or Introduction.

I suggest the comparisons made in the discussion between the study site and other regions can be summarised in a table.

Attention is needed to correct english in the figure legends.

The results are relevant to the aims however the structure of the manuscript can be improved with simplified listing of the objectives.
Experimental design: I believe attention is necessary to describing the methods more clearly. Overall I understand from the manuscript that the research is relevant and adresses an overlooked approach however this can be communicated more clearly.
Validity of the findings: The conclusions are clear, however can be communicated more clearly. I believe more focus should be given to the comparison of the methods for interpolation and less to the description of the community structure at Madagascar reef. Community descriptions can be detailed in supplementary information, but I suggest this is not appropriate for a research publication and is more fitting of a survey report.

It is speculated in the abstract that patchy habitats and community composition "could benefit many mobile species" but this is not supported by findings in the paper.
Additional comments: Abstract
Text although clear could be simplified to focus on the objectives. For example from the sentence starting with “we compare…” at line 20 in the Abstract , text could be modified as follows and to also follow the order of objectives outlined at the end of the introduction:

We compare the accuracy of inverse distance weighting (IDW) and ordinary kriging (OK), to predict the distribution and abundance and identify hotspots of habitat forming organisms on a poorly studied coral reef. Hard corals, octocorals, macroalgae, sponges and zoanthids of Madagascar reef, of the Yucatan continental shelf are important biodiversity and fishery resources.

Followed by a description of the habitat:
“The deeper sandy environments of the leeward and windward regions are dominated by macroalgae and octocorals. However, the shallow rocky
environments of the reef crest have the highest richness of habitat-forming organisms with high abundances of octocorals and macroalgae, and patchy dominance of sponges, Millepora alcicornis and zoanthids, creating high
habitat heterogeneity.”

Then followed by the summary results from comparing IDW and OK…

Introduction:
The manuscript provides suitable content, structure and references in the introduction. I suggest attention is given to improve the text in line with the following examples:

Line 75: can the authors identify the “complex procedures” and/or explain why they are undesirable here.

Lines 76-78: Over use of “it’s” becomes confusing. I suggest:
“However, accuracy of remote sensing diminishes as water turbidity and depth increase light absorption by the water column (Lucas & Goodman, 2014). Remote sensing also has limited ability to identify taxa and accurately estimate abundance (Kutser & Jupp, 2006).”

Lines 84-86: Use of “having” is awkward English and I suggest modification for example as:
However, there are many relatively small coral reefs (e.g. ~1 km2), in deep or turbid environments, which are a conservation priority (Cohen & Foale, 2013).

Lines 111-114:
I suggest the outline of the objectes is reworded to combine i with ii, and iii with iv. For example:
“The present study (i) gathered baseline information of the abundance and community structure of HFO at Madagascar Reef, and (ii) compared the accuracy of IDW and OK to interpolate abundances of HFO and synthesize this information”

Methodology
Line 118: the Sampling design is a little unclear. I believe the explanation can be improved. I suggest clearly stating how many transects were used and how long they were, then how many photo quadrats were taken on each transect, followed by the size of each photoquadrat.

Line 129: I suggest changing the wording to :
“Relative abundance of habitat forming organisms was estimated as percent cover in each photograph…”

Line 139:
I suggest changing “of each photography” to “from each photograph”

Line 142-143
“…are of each other,” please change to “…are to each other…”

Information at lines 143 to 149 is possible more appropriate in the introduction or discussion.

Lines 156-157: the following sentence is unclear: “The parameters of searching window were the same for both methodologies.”

Line 163: in reference to “Millepora alcicornis (millepora)” is this a taxonomic subspecies in backrets or is this to indicate further reference in the text or graphs uses “millepora”

Lines 163-165:
“…the inclusion in the model of scleractinian corals produced overestimations on the
predictions given the small colony sizes found in situ (< 25 cm2).”
This should be discussed in terms of where the methodology is appropriate, for example are small corals common in deep and turbid environments and therefore is this method appropriate for corals?

Results
Line 192: I suggest avoinding language such as “so did”. This can be unclear for the intended meaning. A suggested alternative is: “…the percent cover of sandy substrate and macroalgae increased…”

Legend of Figure 1
Change “Localization” to “Location”

Legend of Figure 6:
Correct “fistribution”

Figure 7:
It is unclear to me the term HFO “richness” is appropriate. The description of how this was calculated at line 169 in the methods indicates it is a presence absence result to show how many types of HFO are present.

Discussion
Line 303: Start sentene with a capital “however,…”

I suggest that the comparison of the community structure of Madagascar reefs is not the key result and therefore should not occupy a large initial section of the discussion. Perhaps this can be summarized in a table in the results? It is currently confusing to follow, partly as a result of the use Endlish language, the multiple locations and organisms and also because the organisms referred to are not always specified. For example Lines 307 to 310 do not specify octocorals.

Lines 318-319:
I suggest rewording similarly to:
“Our study found IDW to be a better methodology than OK for interpolating the abundance of coral reef habitat forming organisms.” However I also suggest the authors discuss the limitations for small corals, as suggested in reference for Lines 163-165

Line 343-344
I suggest changing the wording as underlined: “Our interpolations showed how the space of the reef was occupied by the HFO in a mosaic fashion.”

I suggest the discussion can be started with reference to the current statement at lines 396-399, because this is the main conclusion of the study:
“The only published past studies using IDW in coral reef sessile organisms found this method as a good interpolator for coral cover (Walker et al., 2012; D’Antonio et al., 2016); our results agree with them and we extend its applicability to other important sessile organisms that are emerging under climate change (Norström et al., 2009).”


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: ORDINARY KRIGING VS INVERSE DISTANCE WEIGHTING: SPATIAL INTERPOLATION OF THE SESSILE COMMUNITY OF MADAGASCAR REEF, GULF OF MEXICO
Review round: 2
Reviewer: 1

Output:
Basic reporting: Good basic design and implementation of the study.
Experimental design: Abstract:
This is a good summary of the study

Introduction:
Scleractinian needs to be capitalised (line 47 and elsewhere)

line 63 - just a couple - replace with few

good intro

Methods
make sense

Results
Figure 3 would be better placed in the methods, where the different habitats are introduced

Discussion
terminology of Millepora is still confused, the authors use:
Millepora alcicornis (millepora)
or just the species name or just millepora.
at first mention, the authority is missing.

I suggest using Millepora alcicornis or M alcicornis for consistency.
Validity of the findings: no comment
Additional comments: na


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

Input:
Title: HERMIT CRAB RESPONSE TO A VISUAL THREAT IS SENSITIVE TO LOOMING CUES
Review round: 1
Reviewer: 1

Output:
Basic reporting: Figures 1 and 2 don't contain any scale information. Authors should provide scale.

Figure 3: Authors suggest that the stimuli expanded and contracted exponentially to mimic a predator approach with constant speed (lines 160, 166). This is inconsistent with this figure where the size in pixels is increasing linearly. Authors should clarify the discrepancy and depict the stimulus accurately in this figure.

Figure 5: Authors should indicate on the figure whether these differences are statistically significant.

Authors conducted 15 stimulus presentations for each crab. I suggest that they include the data for each individual crab's probability of hiding and the number of times it took for each crab to hide in the data.

Line 45, 56: Reference to Yilmaz and Meister (Curr Biol. 2013 Oct 21;23(20):2011-5. doi: 10.1016/j.cub.2013.08.015. Epub 2013 Oct 10.) would be appropriate as it is a more recent study than the ones the authors cited.
Experimental design: No comment
Validity of the findings: The authors claim in the abstract that the quicker the stimulus expanded the slower the rate of habituation, however in the results they say (line 223) “because large number of subjects failed to habituate, trials to habituation was not analyzed as a measure of reaction.” I suggest taking this out of the abstract as a main finding.
Additional comments: The authors presented the results using clear language. I suggest the paper will be ready for publication after the modifications stated above.


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: HERMIT CRAB RESPONSE TO A VISUAL THREAT IS SENSITIVE TO LOOMING CUES
Review round: 1
Reviewer: 2

Output:
Basic reporting: The paper is well-written and clear, and appropriate literature is cited. However, the organization of the Methods and Results sections are confusing and seem to overlap, and there seem to be multiple Results and Methods headings. I suggest that the authors have a methods section where they describe the experimental setup, animals, etc., and then a Results section where they describe all the results.
Experimental design: I'm curious as to why the first experiment used a star stimulus, and the second used naturalistic stimuli? That makes it difficult to ask whether the naturalistic stimulus was more effective. Did all the crabs escape in response to the hawk and crab images? If it's not possible to do additional experiments, I would not reject for this, but the data should be included if available.
Validity of the findings: It is not clear from figure 2 whether there was any statistically significant difference in the hide latency for the different expansion rates.
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: BGDMDOCKER: A DOCKER WORKFLOW FOR DATA MINING AND VISUALIZATION OF BACTERIAL PAN-GENOMES AND BIOSYNTHETIC GENE CLUSTERS
Review round: 1
Reviewer: 1

Output:
Basic reporting: The manuscript submitted by Cheng et al. describes BGDM Docker - a tool specifically designed to evaluate bacterial pan-genomes. BGDM Docker integrates a software for genome annotation (Prokka), a software for pan-genome analysis and classification (PanX) and a software dedicated to identify clusters of biosynthetic genes, along with the metabolites that can be produced from such clusters (anti-SMASH). These  software are implemented in a single docker container, along with all dependencies necessary for deployment and use in different computing environments. Overall, BGDM Docker represents an interesting tool for comparative genomics analyses and its implementation in a docker platform helps to facilitate installation and ensure reproducibility of data analyses across laboratories.

The manuscript is sound and adequately written, highlighting several important aspects regarding the currently available tools for pan-genome analysis and the contribution of docker-based systems to the development of bioinformatics.

There are a few points, however, that must be checked by the authors:

-	The software is available for download from the manuscript´s website and also from github. However, it would be interesting if authors also made it available from Dockerhub  (https://hub.docker.com/), since, in this case, the functional docker image would be directly available for testing and download, through docker daemon.

-	Several references, cited along the text, are not present in the manuscript´s reference list.

-	Some links in the manuscript´s webpage do not seem to be working properly (such as http://42.96.173.25:8000/bamf_gbk44, for example).
Experimental design: Authors provide an adequate description of the software capabilities, as well as detailed instructions for its implementation and use. However, these instructions suggest that users implement the three software available in the BGDMdocker container  (Prokka, PanX and anti-SMASH) independently. Wouldn´t it be simpler (particularly to unexperienced users) to integrate them though a single script command (pipeline)?

Moreover, as a suggestion, it would be interesting to provide in the instructions an option to run BGDMdocker without having to enter the container, which should facilitate integration of this software with external pipeline-building tools like Snakemake and Luigi, among others. One option would be to provide the command/script in the Docker CMD option, so the end user can execute the pipeline through a command similar to the following:

$ sudo docker run -it BGDMdocker $pipelinecommand $filein

This command could be called in Snakemake, for example, integrating the entire workflow.
Validity of the findings: BGDMdocker provides interesting results and has advantages when compared to many of the currently existing tools for pan-genome analysis. However, the data provided through the manuscript´s website is difficult to download, since the files ad up to more than 600 Mb each, probably due to the presence of large fasta files, which are not relevant for a reader that only wishes to evaluate the software´s capabilities. Thus, while this is not a major point in this critique, I would recommend that authors provide the more relevant output files, such as images, tables and html pages separately in the github or figshare repositories.
Additional comments: The manuscript describes a worthwhile tool for pan-genome analyses of bacterial genomes. Its implementation in a docker container shall facilitate its distribution, installation and use in different computing environments, contributing to generate reproducible results across laboratories. The manuscript is sound and can be considerably improved with a few modifications, such as the ones described above.


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: BGDMDOCKER: A DOCKER WORKFLOW FOR DATA MINING AND VISUALIZATION OF BACTERIAL PAN-GENOMES AND BIOSYNTHETIC GENE CLUSTERS
Review round: 1
Reviewer: 2

Output:
Basic reporting: The English writing is clear.
More background of Docker should be given.
The data and software of the manuscript is public available.
Experimental design: The research idea is novel, which introduces BDGMdockter to be utilized in biomedical research. Detailed instructions to implement BDGMdockter are also given.
Validity of the findings: No comment. Please check General comments.
Additional comments: This work describes a pipeline “BDGMdockter” for analysis and visualization of bacterial pan-genomes and biosynthetic gene clusters. The “BDGMdockter” could be used in PC and distributed in cloud service, which might be useful in biomedical research.

1. github is also a popular free platform for software repository. What is the advantage of Docker compared to github?
2. BDGMdockter is basically a wrapper for existing software Prokka, panX, and antiSMASH. However, the utilization of BDGMdockter seems complicated for biologists to understand with too many command lines and needs high privileges of computer such as “sudo”. Is there any way to use BDGMdockter without root privilege? It would be nice if more docker background could be introduced and the command lines of BDGMdockter could be simplified.
3. In the RESULT section, the authors claim “Fast and reproducible building of the BGDMdocker workflow across computing platforms using Docker” line 95. However, I did not see how “fast” BGDMdocker is and how “reproducible” BGDMdocker is. Maybe it is a good idea to show the running time and memory need for the software, and produce the results which have been already reported by other literature.
4. Line 126. The author claim “a website was built for the interactive exploration of the B. amyloliquefaciens pan-genome and biosynthetic gene clusters using the BGDMdocker workflow”. However, I did not see how to upload the bacterial sequences, submit the job and produce the Table 1, Figures 2 &3 from the website http://pangenome.zggskj.com/home if the interactive exploration is what it means. A detailed steps should be given for interactive exploration.


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: BGDMDOCKER: A DOCKER WORKFLOW FOR DATA MINING AND VISUALIZATION OF BACTERIAL PAN-GENOMES AND BIOSYNTHETIC GENE CLUSTERS
Review round: 2
Reviewer: 1

Output:
Basic reporting: No comments
Experimental design: No comments
Validity of the findings: No comment
Additional comments: The authors address my 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: ON THE RELATIONSHIP BETWEEN TUMOUR GROWTH RATE AND SURVIVAL IN NON-SMALL CELL LUNG CANCER
Review round: 1
Reviewer: 1

Output:
Basic reporting: The text is well written in clear, professional English to a good standard.
Experimental design: Lines 133 - 140. This section would be improved if something could be said about these three trials. For example, were these trials in a primary or metastatic setting? Were patients previously treated? While there are references to the trials, it would aid the reader to have at least some basic background without having to look up three different references.

The trial referred to as Paclitaxel/Carboplatin (ref 23) does not appear to have used Paclitaxel - the trial was actually comparing pemetrexed-carboplatin with etoposide-carboplatin. The reference to Paclitaxel/Carboplatin is made in several places in the text and should be corrected throughout.

Additionally, there is no discussion of how these trials were selected. Were they the only three trials with data availability for the model, or were these three selected from a pool of candidate studies? If selected from a pool what were the selection criteria?
Validity of the findings: No comment.
Additional comments: An interesting question for the discussion is whether disease setting has an impact on these analyses? For example the pemetrexed/carboplatin trial was in chemo-naive patients, whereas the other two trials were in previously treated patients. Does it make sense to imagine that the disease trajectory would be the same in these two very different populations? Is it correct to assume that the same model would apply? Similarly, the point is made in the discussion that these results pertain to NSCLC - some discussion on whether these results generalise to other solid tumours would be helpful.


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: ON THE RELATIONSHIP BETWEEN TUMOUR GROWTH RATE AND SURVIVAL IN NON-SMALL CELL LUNG CANCER
Review round: 1
Reviewer: 2

Output:
Basic reporting: I found this to be a very interesting and well-written paper with appropriate references to the relevant literature. The paper was well-structured and cogently argued.
Experimental design: The paper addresses a very important question - how to predict phase III outcome for a
new anti-cancer treatment using early clinical data.

The statistical analyses carried out on the data (tumour re-growth rate, time to tumour re-growth and survival) were entirely appropriate and well-presented.
Validity of the findings: I found the findings compelling and therefore the conclusions of the statistical analysis are very important and have important implications for future design of phase III clinical trials for anti-cancer drugs.
Additional comments: This is a very sound paper with interesting findings which deserves to be published.

Some minor typographical errors which should be corrected are:

lines 51, 52: "the evaluation of a drugs’ efficacy..." -> "the evaluation of a drug's efficacy..."

line 52: "The drugs effect on target lesions are reported..." -> "The drug's effect on target lesions is reported..."

line 199: "Details of the patients imaging and survival characteristics..." -> "Details of the patients' imaging and survival characteristics..."

line 221: "it’s clear that..." -> "it is clear that..."

line 240, 241: "However, by re-phrasing the question to, do TTG and GR correlate to survival post progression, to..." ->  "However, by re-phrasing the question to, do TTG and GR correlate to survival post progression?, to..."


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: ON THE RELATIONSHIP BETWEEN TUMOUR GROWTH RATE AND SURVIVAL IN NON-SMALL CELL LUNG CANCER
Review round: 2
Reviewer: 1

Output:
Basic reporting: Pass.
Experimental design: Pass.
Validity of the findings: Pass.
Additional comments: Thank you for addressing the various comments raised in the previous round. I am happy for this interesting paper to be published in its revised form.


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: ON THE RELATIONSHIP BETWEEN TUMOUR GROWTH RATE AND SURVIVAL IN NON-SMALL CELL LUNG CANCER
Review round: 2
Reviewer: 2

Output:
Basic reporting: All good.
Experimental design: All good.
Validity of the findings: All good.
Additional comments: All requested changes were made - the manuscript is ready for publication in my opinion.

Overall, a very timely, original and interesting piece of work.


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

Input:
Title: TRANSCRIPTOMIC PROFILING OF MTOR AND RYANODINE RECEPTOR SIGNALING MOLECULES IN DEVELOPING ZEBRAFISH IN THE ABSENCE AND PRESENCE OF PCB 95
Review round: 1
Reviewer: 1

Output:
Basic reporting: Well-written paper, but some of the figures are not necessary or could be moved to supplemental information.  Raw data were not shared as supplemental information.  Discussion is too long given the limited scope of the study.
Experimental design: Solid study but very limited in scope, as the authors only evaluated baseline expression profiles of a handful of genes at three stages of embryonic/larval development.
Validity of the findings: Findings are valid, but the authors should just present relative expression or absolute transcript numbers (not both).
Additional comments: Line 11: Please spell out mTOR upon first use.

Lines 60-61: Recommend moving Figures 1 and 2 to supplemental information.

Lines 230-343: The Discussion is way too long (~5 pages), especially given the limited scope of this study.  Therefore, the Discussion could easily be trimmed down to no more than 2.5-3 pages.

Lines 262-262: While this is true, it's unknown whether chemical uptake into the yolk sac interferes with nutrient availability and utilization during embryonic development.  Please revise this sentence to address this possibility.

Tables 1 and 2: Move to supplemental information.

Figures 3 and 4: These two figures could easily be combined into a single heat-map or volcano plot.

Figure 5: I'm not convinced that these data add value in addition to Figures 3 and 4.  Why not just present as a fold-change or absolute copy number?  Both are not necessary, as these figures rely on the same qPCR data.

Figure 6: Please include images for sense ryr3 RNA probes (negative controls) in order to account for potential non-specific hybridization in situ.


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: TRANSCRIPTOMIC PROFILING OF MTOR AND RYANODINE RECEPTOR SIGNALING MOLECULES IN DEVELOPING ZEBRAFISH IN THE ABSENCE AND PRESENCE OF PCB 95
Review round: 1
Reviewer: 2

Output:
Basic reporting: The article is well written and the background for the study is sufficient.  The figures and tables are professional.   There are no hypotheses stated as this is an observational study of gene expression during ontogeny of zebrafish.
Experimental design: In general the experimental design section is well written.  There is a question about how the RNA check gel was performed and it would be good to have included a picture for the quality of the RNA.
Validity of the findings: The findings are valid but the study is observational.  There was no experiment included.
Additional comments: In this manuscript, the authors map gene expression of key transcripts in the mTOR and RyR signaling pathways in wild type zebrafish embryos at three times points during development.  The genes that they selected are excellent genes for these pathways and they provide primer pairs for the genes of interest.   While it is interesting to know the timing of expression of these genes, I found the study to be descriptive and without an experimental hypothesis.   Apparently this study is setting the baseline for future studies in which they will examine changes in these genes after exposure to contaminants.  In my opinion, the study would be improved if the authors incorporated an actual experiment.

Comments:
1.	The authors extract the total RNA from a pooled set of whole embryos for a set of genes that might give information for brain development.   Since these genes are likely expressed differentially in tissues during organogenesis, they dilute their signal by using whole embryos.  This is a major limitation of the study as the authors point out in line 292.  They might get better results if they could micro-dissect out the various tissues of interest.
2.	How closely were the fish embryos that were pooled synchronized for development?  Since development is rapid, a half hour difference for pooled embryos could erase a significant change in gene expression.
3.	Line 207 – absolute transcript copy numbers --   I have an issue with this.  The authors make cDNA for the amplified segment and purify it and measure the quantity by nanodrop.   However, when doing PCR from a transcript, there is the step of reverse transcriptase that is not necessarily 100% efficient.  If they want to have absolute copy number, they should start with RNA.  They could get RNA for each of their genes by adding a T3 RNA polymerase promoter to one end and transcribing the RNA and quantifying that product before starting the PCR part.   Also it is not clear if the absolute transcript copy number is per embryo or per ug total RNA or???   It would probably be most interesting to have the calculation be per embryo.
4.	Line 43-45 – statement that the two pathways are critical for the development of the nervous system needs a reference.  It seems to me that mTOR would be critical for the whole organism as would the RyR signaling pathways since they both play critical roles in homeostasis.
5.	Line 114.   Was total RNA integrity measured on a denaturing agarose gel – for example MOPS gel?  It might be good to provide an example of the gel since RIN numbers are not provided.
6.	Line 186 – in the text the gene is called “rictora” but in the figures it is referred to as “rictor”   Please correct and make them the same.
7.	 Line 228.   I don’t think that it is possible to quantitatively compare the staining for in situ across the different embryos shown in Figure 6.  The magnification seems to be different for the embryos and there should be a standard against which to measure the intensity of the stain for quantitation.   The statement  …. “even though expression throughout the rest of the body is still low…”  doesn’t seem to match the very dark staining seen in the rest of the body in the other panels at earlier stages without appearance in the brain.
8.	Line 249 --   I don’t think the authors can comment on whether the increased expression of rictora and rptor promote mTOR activity since (a) they have not measured mTOR activity and (b) mTOR activity is promoted at the level of phosphorylation
9.	Line 253 – Chemical perturbations may change the expression patterns for these genes, but the authors have not performed these experiments.  If they have, they should include them in the paper as it would strengthen the paper.
10.	Line 263 – not all yolk sacs are nutritionally equivalent  and provide the same amount of nutrients.  There are some eggs of lower quality that may have inferior yolk composition, especially if mom was exposed to contaminants.
11.	Lines 307-310.   The authors have not shown in situ for Ryr1 or Ryr2 and thus cannot comment on spatial expression in the brain in zebrafish in their experiment.  They quote other authors, suggesting that RyR spatial expression is already known for zebrafish and thus takes away the novelty for this study.

Minor comments
Line 47   delete “the”   should read …throughout lifespan….
Line 91  Add “was”   should read….and RNA was extracted…..
Line 246   rictora or rictor?


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: TRANSCRIPTOMIC PROFILING OF MTOR AND RYANODINE RECEPTOR SIGNALING MOLECULES IN DEVELOPING ZEBRAFISH IN THE ABSENCE AND PRESENCE OF PCB 95
Review round: 2
Reviewer: 1

Output:
Basic reporting: The authors have adequately addressed all previous comments.
Experimental design: The authors have adequately addressed all previous comments.
Validity of the findings: The authors have adequately addressed all previous comments.
Additional comments: Recommend accepting for publication.


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

Input:
Title: AGE-BASED AND REPRODUCTIVE BIOLOGY OF THE PACIFIC LONGNOSE PARROTFISH HIPPOSCARUS LONGICEPS FROM GUAM
Review round: 1
Reviewer: 1

Output:
Basic reporting: No comment
Experimental design: No comment
Validity of the findings: No comment
Additional comments: The paper ‘Age-based and reproductive biology of the Pacific longnose parrotfish Hipposcarus longiceps from Guam’ by Taylor and Cruz provides important demographic information about a commercially important species. The manuscript is written very well. The Introduction provides an excellent coverage of the background information and sets up the reasons for this study in the context of information gaps. The Methods and Results were explained well and, despite being unfamiliar with a couple of the analyses performed, literature support was provided to justify their use. I particularly liked the clear structure of the Discussion, which clearly outlined the importance of the findings in the context of relevant literature. Thank you to the authors for providing a manuscript that was enjoyable to read.

I have very few suggestions for improving this manuscript. However, the comments listed below should be considered…

Abstract:
(i)	Line 20 – “… and tend to change sex to males…” This is rather ambiguous. I suggest rewording this.

Introduction:
(i)	Lines 43-44 (and elsewhere) – The common name for this species is stated in the title so I do not see a reason for it to be included frequently throughout the manuscript. All references to the common name should be changed to the scientific name. This is the second time the common name for this species is mentioned in this paragraph.
(ii)	Lines 45-46 – What is meant by “… among the larger species that occurs in Guam…”? Larger species of what? Of parrotfishes?
(iii)	Lines 55-56 – “… as can be confirmed histologically.” This needs to be referenced.
(iv)	Lines 60-61 – What is meant by “…whereby age at female maturation had the strongest relationship to…”? Does this mean they mature earlier?
(v)	Lines 62-63 – Similar to my comment above, what does “… with corresponding life-history traits generally at the far-end of the spectrum.” actually mean? Does this mean at the lower/higher ends of the spectrum or at both ends? Also, what life-history traits are actually getting referred to here? Is it age/size at maturation and sex change?
(vi)	Lines 66-67 – The way this sentence is constructed it implies that Taylor (2014) suggests that the Guam populations will be larger-bodied and longer-lived. Is this actually the case? If so, it may be more appropriate to place this reference at the end of the sentence. If not, then an additional reference is needed at the end of the paragraph.

Methods:
(i)	Line 133 – There appears to be an issue with the insertion of a symbol here with f() being in a dashed, square box.

Results:
(i)	Line 209 – What is meant by ‘slightly invariable’?

Discussion:
(i)	Lines 272 & 356 –Why is the common name mentioned here and not the scientific name? (see earlier comment)

Conclusion:
(i)	Lines 358-360 – I find this final sentence of the Conclusion a little confusing in what was otherwise a well-written and clear manuscript. I suggest clarifying this sentence to strengthen the ‘take-away’ message from this study.

Figures:
(i)	Figure 2 (caption) – There is repetition of words in this caption that is not needed. I suggest shortening it to read “(a) Sex-specific length- and (b) age-frequency distributions of sampled Hipposcarus longiceps from Guam.”


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: AGE-BASED AND REPRODUCTIVE BIOLOGY OF THE PACIFIC LONGNOSE PARROTFISH HIPPOSCARUS LONGICEPS FROM GUAM
Review round: 1
Reviewer: 2

Output:
Basic reporting: The quality of the manuscript is so high, so I gave only a handful suggesting and comments.
Experimental design: The research question was well defined, and perfect for the journal.
Validity of the findings: Data is enough for this large sized species, and all result are well reliable.
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: FACILITATIVE AND COMPETITIVE INTERACTION COMPONENTS AMONG NEW ENGLAND SALT MARSH PLANTS
Review round: 1
Reviewer: 1

Output:
Basic reporting: This paper describes a well-designed experiment that tests the degree to which plant-plant interactions are positive and negative across the life history of a perennial forb and determines the redundancy of three matrix species. The analyses are appropriate. The writing is clear and professional. The authors cite the relevant literature both within and outside their study system to support their approach.

To prepare the reader for the methods, the authors should be more specific in the introduction regarding the meaning of light environment and salinity. They should also be more detailed about the importance of (reasoning behind) the 'mimic' treatment.

Specific comments for the introduction:
Line 81 an e.g. or example is needed here

Line 89 examples of species specific factors that preclude functional redundancy should be included here.
Experimental design: no comment
Validity of the findings: The authors are careful in interpreting their results, but so much so that there is little that those working outside the salt marsh system can take away from the paper. The authors should add context to their conclusions: how do these results apply to other ecological systems? Improving the text supporting the light and the salinity measurements will help with this.
Additional comments: none


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

Input:
Title: FACILITATIVE AND COMPETITIVE INTERACTION COMPONENTS AMONG NEW ENGLAND SALT MARSH PLANTS
Review round: 1
Reviewer: 2

Output:
Basic reporting: ABSTRACT:
Lines 27, 29: “Three matrix-forming grass and rush species” implies three of each. Use instead, “one species of rush and two grass species” for clarity.

Line 31: Suggest changing “forbs” to simply the species name Aster tenuifolius since this was the only forb analyzed in the experiment, and “forbs” implies more than one forb species.

Line 33-35: “There was no statistically significant variation among matrix species
in their net or component effects, however, the competitive effect of J. gerardi was
negligible, especially compared to that of D. spicata.” Unclear wording.

Line 34: “…net or component effects. However, …”

INTRO:

Line 50: Although elucidating facilitation within your study system is the main aim of the research, this first line should be inclusive of both general types of interaction (facilitation and competition) as the concept is introduced initially. I.e. “Research on community organization in ecology has long recognized that species interactions are often composed…”.  Then, honing that down through this first paragraph to the last lines,

Lines 63-65: Try “The prevalence of facilitation in instances of environmental stress or particular life-history stages does not minimize the role played by competition…” Omit “trivialize”, I’m not sure any ecologist would ever consider competition a ‘trivial’ component of a community, but I think I get at your point.

Line 99: ”…may not, leading to a de-coupling…”

Lines 107-108: Same comment as above in abstract regarding clarity of grasses and a single rush species. Also, comma unnecessary after “rushes”

METHODS
Line 113-114: It might be good to include the nearest town or other geographical landmark (or Lat/Long?) to specifically identify Nag Creek Marsh. Being unfamiliar with the area, it took a bit of time to locate the marsh online.

Line 143: You state that A. tenuifolius both is a perennial and that it germinates in early spring. An individual of A. tenuifolius from a previous season is not ‘germinating’, but rather re-emerging from established below-ground rootstock. Is there a difference in emergence/germination times?

DISCUSSION
Line 265: “…standing dead biomass, …”
Experimental design: Line162-163: “Shade cloth…in the field.” The white painting may have provided elevated photosynthesis (over either darker bare substrate or surrounding vegetative matrix). How was this either accounted for or monitored?

Lines 170-171: Were the dug J. gerardii plots replaced in the exact same hole from which they were dug, or were they moved to one of the four plots within the block to simulate hole size or local substrate mismatch likely part of transplanting the other matrix species?

Lines 188-189: is this growth timeline (June to August) the full breadth of seasonal growth of these species? There needs to be more life-history information within the Introduction, or here, to show why these times were chosen. The Introduction identifies “early spring” as the germination time (and see my previous comment regarding perennials), and August seems early for the end of growth (thinking more September/October).
Validity of the findings: No Comment.
Additional comments: It is noted that this research was done in the summer of 1999. The majority of the references for this work are 15 or more years old, with a few exceptions. Before publication, I strongly recommend including more current research on the topics included, with attendant descriptions of how this work remains novel and contributory within that context.


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

Input:
Title: DNA BARCODING REVEALS THE MISLABELING OF FISH IN A POPULAR TOURIST DESTINATION IN BRAZIL
Review round: 1
Reviewer: 1

Output:
Basic reporting: The main issue I had with this paper was its English style. While understandable, it has many errors in grammar and usage. I suggest the authors firstly revise the paper (after considering more specific comments made below) and then get this revised version carefully read and corrected either by a native English speaker or by a professional editing company (there are on-line companies that do this).

The study itself I found to be interesting and satisfactorily carried out, and is within the scope of PeerJ. Raw data, in the form of the barcode sequences, are supplied.
Experimental design: Analytical methods, both the initial barcoding protocols and subsequent assignment of species, were appropriate and generally satisfactorily described.
Validity of the findings: The results and findings were interesting and adequately referenced. Conclusions were well supported by the data.
Additional comments: 1.	The major recommendation – apart from improving its English style – is to reconsider and improve the layout of the two Tables. In my view these can be considerably improved by:
a.	A: Re-arranging the numbering, so that the correct assignations form Table 1 and the incorrect assignations Table 2. This will obviously then require some small textual changes too.
b.	B: Subdividing each Table into two subsections, one being the fishmongers and the other the Japanese restaurants.
c.	C: Spell out the names of the genera – as there are many of them, it’s not sufficient to identify them here simply by initial letters.
d.	D: Aren’t the numbers in parentheses the best or highest match comparisons, in percent, not the lowest match comparisons?

2.	Lines 29-33. Explanation of the Cat by Hare Project is not readily intelligible as given – please clarify.

3.	Line 30. Lahiri and Nurnberger, not K. and Nurnberger.

4.	Line 89. Blast and BOLD respectively, not BOLD and Blast respectively.

5.	Lines 96-103. Please explain the relevance of giving BOLD BIN numbers for these species (or omit them).

6.	Line 133-134. Genus not genre.

7.	Please check spellings of all scientific names in text and tables. I noticed misspellings of L. niloticus (Table 2) and line 162 O. kisutch. Also give all genus names in full in the text, as you are dealing with a wide variety of genera, not just one or two.

8.	Check references. DNA should be given as DNA not as Dna. Line 274 – delete d. Some journals also have part numbers given – e.g. (1) in Barbuto et al. Is this necessary? I suspect these should all be deleted.


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

Input:
Title: DNA BARCODING REVEALS THE MISLABELING OF FISH IN A POPULAR TOURIST DESTINATION IN BRAZIL
Review round: 1
Reviewer: 2

Output:
Basic reporting: The results are described in a way that is not always easy to follow and the language is confusing at some points. An editing by an English native speaker or an editing service is highly recommended.

The introduction should include a brief description of the DNA-based methods used in species identification. It should describe the concept of DNA barcode and its advantages and disadvantages. For example:
L. Wilson-Wilde, J. Norman, J. Robertson, S. Sarre, A. Georges, Current issues in species identi?cation for forensic science and the validity of using the cytochrome oxidase I (COI) gene, Forensic Science, Medicine, and Pathology 6 (2010) 233–241.

The authors should mention in the Introduction section the previous initiatives for the implementation of forensic standards in the identification of species, particularly in the field of food forensics. For example:
F. Teletchea, C. Maudet, C. Hänni, Food and forensic molecular identiﬁcation: Update  and challenges, Trends in Biotechnology 23 (2005) 359–366.
M. Woolfe, S. Primrose, Food forensics: using DNA technology to combate misdescription  and fraud, TRENDS in Biotechnology 22 (2004) 222–226.
Experimental design: The methods should be better described.

Which type of material and conditions were implemented for the sample collection? Sterile material? I’m worried about the possible contamination of the samples during the tissue collection. The authors should describe in more detail the sampling process and discuss possible contamination issues. For example, the sample fish piece might have been in contact with pieces from other species, and a cross-contamination is possible.

The sequencing reaction should be briefly explained. Which reaction conditions were used? Which primers? Do the authors sequence the PCR products in both directions?
The parameters used in the Blast and BOLD searches should be indicated (blast algorithm, database, etc). The experiment has to be described in a way that anyone could replicate it.

Lines 92-103. The paragraph is not clear. Why do they used the IUCN Red List of Threatened Species status? Which species are listed here?
Lines 105-108: The English should be revised.
Line 35: parenthesis are missing in the reference
Lines 48-48: the sentence should not be in italics
Line 72: PCR reaction is redundant. The R is for reaction.
Lines 83-85: The sentence should be revised. The meaning is not clear.
Line 172: it should be “species”
Validity of the findings: The approach used by the author is valid as it has been used by others before.
Additional comments: no comment


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

Input:
Title: DNA BARCODING REVEALS THE MISLABELING OF FISH IN A POPULAR TOURIST DESTINATION IN BRAZIL
Review round: 2
Reviewer: 1

Output:
Basic reporting: I found the results of this study were presented much more clearly than in the original submission, but the English style remains, for me, poor. In fact I find it hard to believe that two ‘professionals’ had separately read the manuscript and all their comments were incorporated – they cannot have been highly proficient in the English language! The title of the paper is now greatly improved, but the wording of the Abstract (for example) remains virtually identical to the original Abstract. It is true that the text can, with a little difficulty, be understood by an English speaker, but in my view it still needs improvement prior to publication.

I continue to recommend editing by a professional English speaker or editing service.
Experimental design: Acceptable
Validity of the findings: Acceptable
Additional comments: Some specific points:

1.	Line 18. ‘30% of fraud in fisheries’ Isn’t this really 30% mislabelling in fisheries? Later in the Abstract the authors mention that not all cases of mislabelling are necessarily fraudulent.
2.	Lines 52-58. I did not find the short explanation of DNA barcoding to be useful, and could even be confusing to newcomers to the topic.  What is meant by a ‘global system of different mutational rates’? The implication that nucleotides mutate randomly so that 4 * 15 sites would generate 1bn new codes is a considerable simplification of reality – most sites are conserved! Please clarify this paragraph.
3.	Line 79. e to and
4.	Line 80-82. I couldn’t understand this sentence
5.	There are still some spelling errors in scientific names, e.g. for tilapia in Table 1 (but OK in Table 2) and for coho salmon (line 172)
6.	Line 115. What is ‘GL’ – is it degrees of freedom?
7.	Line 169 autosomal
8.	Line 183 what is ‘abastance’?


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

Input:
Title: DNA BARCODING REVEALS THE MISLABELING OF FISH IN A POPULAR TOURIST DESTINATION IN BRAZIL
Review round: 2
Reviewer: 2

Output:
Basic reporting: No further comments to the manuscript.
Experimental design: No further comments to the manuscript.
Validity of the findings: No further comments to the manuscript.
Additional comments: No further comments to the manuscript.


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

Input:
Title: DNA BARCODING REVEALS THE MISLABELING OF FISH IN A POPULAR TOURIST DESTINATION IN BRAZIL
Review round: 3
Reviewer: 1

Output:
Basic reporting: English, and the discussion of the barcode approach at the end of the Introduction, is improved and adequate for publication.
Experimental design: Satisfactory
Validity of the findings: Good
Additional comments: None


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

Input:
Title: EVALUATION OF POTENTIAL REFERENCE GENES FOR REAL-TIME QPCR ANALYSIS IN A BIPARENTAL BEETLE, LETHRUS APTERUS (COLEOPTERA: GEOTRUPIDAE)
Review round: 1
Reviewer: 1

Output:
Basic reporting: The ms is clear, well written in professional English and conforms to professional standards of expression.
Literature references basically cover sufficiently the field context. I would suggest to add the following paper and to accomplish the checklist provided in it.
The MIQE Guidelines: Minimum Information for Publication of Quantitative Real-Time PCR Experiments, by Stephen A. Bustin et al., Clinical Chemistry Apr 2009, 55 (4) 611-622; DOI: 10.1373/clinchem.2008.112797.
The ms includes sufficient introduction and background and demonstrates how the work fits into the broader field of knowledge.
Figures are relevant to the content of the article and appropriately described and labeled. Resolution of Figure 1 is probably too poor. It is suitable for review, but it would be necessary improve the quality in final version. A comment to figure 1 is missing in result sections. Figure 1 indeed shows that generally thorax samples showed higher Cts than head, especially in males and that generally there were no strong differences between male and female samples. A statistical analysis of these differences (ANOVA on different samples for each genes) could improve this statement and Figure 1 as well.
A different table organization is needed to improve ms readability. Tables 1 and 2 could be merged, and Authors should add qPCR parameters for different primer pairs that are now missing (Efficiency, R2, melting temperature). Table 5 and 6 should be provided as supplementary data, since are not crucial. On the other hand, data about different ranking obtained by reffinder on different tissue and different sex are missing. Authors should include new table with reffinder results for different subset of samples analysed, especially to facilitate reading of lines 251-258 in results and 296-300 in discussion. These paragraphs describe key findings of ms, but they need to be rephrased.
The ms is ‘self-contained’ and represents an appropriate ‘unit of publication’.
Experimental design: Research question is well defined, relevant and meaningful, as author stated in discussion lines 284-287.
The investigation was rigorous and performed to a high technical standard.

Methods are generally well described. Nevertheless, the different subset of samples analysed should be clearly stated. In other words, if I understood well, authors run different gene stability analysis to establish reference genes on different sample subsets: i) all samples, irrespective of tissue and sex; ii) all females, irrespective of tissue; iii) all males, irrespective of tissue; iv) all thorax samples, irrespective of sex; v) all head samples, irrespective of sex; vi) female thorax samples; vii) male thorax samples; viii) female head samples; ix) male head samples. Authors should better explain this experimental plan of analyses and including a new table at least including reffinder results for all these analyses.
Validity of the findings: Presented results are statistically sound, but some details on different subset of analysis are missing, as previously indicated.
qPCR Efficiency must be calculated for all primer pairs. Indeed in relative quantity calculation it should be included to obtain more accurate data.
Conclusion are well stated, linked to original research question and limited to supporting results.
Additional comments: Detailed comments not included in above suggestions are:
-In the abstract is missing the fact that L7A and RP18 are also the most stable genes considering all samples irrespective of tissues and sex.
- “Lethrus apterus”, “reference gene” and “RT-qPCR” keywords are redundant since already present in the title.
-Lines 58-42: please rephrase.
- Line 84: Change it, since, reference genes must be evaluated in different specific sample types and treatments. The concept of “universally applicable reference genes” is not reasonable.
- Line 89: change in “the expression level of hormone regulating genes involved in parental care”
- Line 141-142: please add details about negative controls of no-retrotranscribed samples to exclude genomic DNA contamination.
-Line 148 149. Change with the “lowest Ct value” instead of the “highest”. (the lowest Ct value corresponds to the highest cDNA starting quantity)
-Caption to Table 5: rephrase the subset of samples analysed.


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: EVALUATION OF POTENTIAL REFERENCE GENES FOR REAL-TIME QPCR ANALYSIS IN A BIPARENTAL BEETLE, LETHRUS APTERUS (COLEOPTERA: GEOTRUPIDAE)
Review round: 1
Reviewer: 2

Output:
Basic reporting: No comment
Experimental design: 1.	In line 116, the author quoted “Using a draft genome of Lethrus apterus (Rácz et al., 2015)”. Looking at the paper, its not a genome paper, it’s a paper identifying microsatellite markers. Can the author clarify this? Is there a genome of Lethrus apterus?
2.	The amplicon length of some of the genes are quite big, up to 426bp for a qPCR experiment. Its usually recommended to keep amplicon size small and if possible about the same length across all test genes. Is there any particular reason why some of these genes have big amplicons?
3.	The authors mention about sample collection in line 94-96. Was the sample pooled? If so, which samples were pooled?
4.	The number of variables are quite limited, only head and thorax of male and female collection. Although it is understandable that the sample is an environmental sample, but in this type of experiment, it is important to test the candidate genes in different variability (e.g. different stages of development) to measure its stability.
Validity of the findings: 5.	For the gene finder and BestKeeper analysis in Table 4 and 6.  The results shown are for which samples? As there are 4 samples chosen by the authors (head and thorax of male and female), why is there only 1 set of data?  Can the authors comment to make this clearer.
6.	Same goes to NormFinder data in Table 5, why is there only Head and Thorax but there is not male and female data?
7.	Did the authors sequence the PCR products to see if the correct genes were amplified?
8.	Could the author explain why the housekeeping gene was not tested/validated using another gene?
Additional comments: The work presented in this manuscript describe the use of potential reference genes for the use of qPCR based studies in Lethrus apterus. The authors choose 11 candidate genes to test its suitability to become the housekeeping genes.
The data presented appeared to be technically sound for the most part. Most papers are using geNorm, NormFinder, BestKeeper and RefFinder to evaluate the suitability of these candidate genes.


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: DIAGNOSTIC VALUE OF SERUM PROCALCITONIN, LACTATE, AND HIGH-SENSITIVITY C-REACTIVE PROTEIN FOR PREDICTING BACTEREMIA IN ADULT PATIENTS IN THE EMERGENCY DEPARTMENT
Review round: 1
Reviewer: 1

Output:
Basic reporting: 1.	Title: the title is misleading. The authors have assessed the diagnostic value of several biomarkers commonly used in predicting positive blood cultures, not diagnosing sepsis. The title should reflect this.
2.	Abstract: the authors state the “lactate added to procalcitonin helps to improve the performance to differentiate whether blood culture will be positive with either Gram-positive or Gram-negative pathogen”. However, the results presented do not support this conclusion as no biomarker levels are presented. For example, if the patient has elevated lactate and elevated PCT, this information will not help the clinician to differentiate between Gram-positive or Gram-negative bacteraemia. It is rather just an indicator that the patient is more likely have a positive blood culture.
3.	Discussion: too few references are used in the Discussion (only three). The results presented in this paper must be considered in context of other people’s work.
4.	Abstract: abbreviations should not be used in the abstract.
5.	Introduction: the introduction need more detail and relevant references. For example, the previously published results for PCT as a biomarker have been contradictory, which is not mentioned in the introduction.
6.	Introduction: references are missing for several claims. For example, line 79-80; line 86-87. Besides, many of the references are relatively old – there have been a lot of publications in this area the latest years.
7.	Material and method: no information whether one-sided or two-sided test were used.
8.	Material and method: Line 152: the sentence “The p-value in our analysis was set at 0.05” should be reformulated as one cannot set a p-value. Before you perform a test, you decide the significance level (alpha) which is usually set to 0.05.I suggest that you write: “The significance level was set at 5% (p < 0.05).” or “A two-sided p value of <0.05 was considered statistically significant.”
9.	Regarding the number of decimals used for reporting p-values both in text and tables: too many decimals used, I suggest that you use two decimals for p-values >0.01; three decimals for p-values between 0.01-0.001; p-values below 0.001 should be reported as p<0.001.
10.	Regarding the number of decimals used for reporting the AUROC values: too many decimals used, I suggest that you use two decimals.
11.	The 95% CI for AUROC should also be reported, in the text as well as in the figure legends.
12.	Results: Interpretations of results and conclusions are presented in the Result section; this belongs to the Discussion section. For example, line 206: “With regards to the single test item, PCT is still a better choice.” and line 209: “Therefore, a combination of PCT or lactate increases is better against blood culture.”, etc.
13.	Results: all paragraphs starts with “Table X shows…”.  I suggest that the authors report the key results in the text and then referring to the figures and tables in the text.
14.	Sometimes the authors use the term odds ratio (OR) and sometimes diagnostic odds ratios (DOR). This is confusing. The authors should stay put to use one of the terms.
15.	Table 3: cut-off for lactate is written as 19.8 mg/dl? Confusing as the authors have stated 1.98 mg/dl as cut-off elsewhere in the paper.
16.	Table 7: all abbreviations used in the table is not explained in full (e.g., Y, ED, PCT).
17.	Table 7: complete references are missing; only the name of the first author is provided.
Experimental design: 1.	Statistical comparisons between the ROC curves are missing. In order to determine whether there is a significant difference or not you must test for that, not only rank them in order. I suggest that you use DeLong’s test for comparison of two correlated ROC curves (DeLong et al. Comparing the areas under two or more correlated receiver operating characteristic curves: a nonparametric approach. Biometrics. 1988;44(3):837-45).
2.	The reasons for selection of the cut-off values for lactate, PCT and CRP should be described. For example, why cut-off values for lactate >1.98 mg/dL and CRP>0.8 mg/dl? These cut-off values have not been used in previous sepsis studies.
3.	Material and method: Line 127: information about the blood volume drawn from each patients is missing as well as the number of blood culture bottles incubated for each patient.
4.	Material and method: neither information nor reference are given for which bacteria species were interpreted as pathogenic or contaminant.
Validity of the findings: There are many caveats in this paper which limits it validity.
1.	Definition of sepsis: this is my major concern about this paper. For several reasons, the results presented in this paper cannot be used in diagnosis of sepsis as claimed, only to predict positive blood cultures. Firstly, I would like to stress that positive blood culture is not the same as sepsis. Many patients suffer from sepsis without having a positive blood culture (only 30-40% of the septic patients have a positive blood culture) and vice versa, many patients can have a positive blood culture without having sepsis. Besides, sepsis may be caused by other pathogens than bacteria. The previous definition (Sepsis-2) of sepsis included proven or suspected infection accompanied by systemic inflammatory response syndrome (SIRS), whereas according to the new sepsis definition (Sepsis-3), sepsis should be defined as life-threatening organ dysfunction caused by a dysregulated host response to infection (Singer et al., 2016. The Third International Consensus Definitions for Sepsis and Septic Shock (Sepsis-3). JAMA 801-810:(8)315). Secondly, the patients enrolled in the study were not selected based on a clinical suspicion of sepsis. Patients were enrolled if all four tests (blood culture, PCT, hs-CRP, lactate) were performed within 24 hours after admission to the hospital – that´s a huge different. No information is provided about the clinical diagnosis nor the reason for taking blood cultures.
2.	The paper studied the predictability of positive blood cultures in adult patients, not sepsis. This must be made clear throughout the whole paper.
3.	Cut-off values: the selected cut-off values have not been used previous sepsis studies and are in disagreement with the ones generally recommended for clinical practices.
4.	The levels for the different biomarkers should be presented as well in the Result section.
5.	Sometimes the authors write that they have studied blood cultures positive with Gram-negative bacilli or bacteria/Gram-positive cocci, and sometimes blood cultures positive with Gram-negative /Gram-positive pathogens. This inconsistency is confusing – what have actually been studied? How did the authors evaluate findings like Gram-positive bacilli? Where they considered as negative or contaminations or excluded from the study?
6.	Raw data file is supplied, that´s excellent. However, no information is given about the numbers of positive blood culture bottles, no either is any information about the findings classified as contaminants. As the number of positive blood cultures are of importance for deciding whether it’s a pathogen and contaminant it should be provided. What about polymicrobial findings? What about fungal findings? The raw data file also contains some Chinese characters not possible for me to understand. Moreover, units for the biomarkers measurement are missing.
7.	Accuracy is another useful performance measure, not included in the paper. It would have been a nice addition to the paper.
Additional comments: 1.	Discussion: the studies you compare with have used other cut-off values which makes it difficult to compare. It is more correct to only compare results between studies using same cut-off values.
2.	The use of abbreviations: abbreviations should generally be avoided. For the first reference to a term in the text, the term should be used in full with the abbreviation included in brackets. For the remainder of the text the abbreviation should be used. Moreover, I recommend that the authors only used standard abbreviations commonly used (e.g., DOR, CI, PCT, hs-CRP, ROC).
3.	Line 238: sepsis is NOT defined as a bloodstream infection.
4.	Line 242: “In this study, PCT was compared with conventional competing tests, such as hs-CRP or lactate,…”. I guess that “or” should be changed to “and”?
5.	Line 298: the authors write that PCT has a better diagnostic ability for GNB, but in comparison to what?


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: DIAGNOSTIC VALUE OF SERUM PROCALCITONIN, LACTATE, AND HIGH-SENSITIVITY C-REACTIVE PROTEIN FOR PREDICTING BACTEREMIA IN ADULT PATIENTS IN THE EMERGENCY DEPARTMENT
Review round: 1
Reviewer: 2

Output:
Basic reporting: See general comments
Experimental design: See general comments
Validity of the findings: See general comments
Additional comments: Thank you for asking to review this paper. This retrospective analysis assessed the utility of lactate, PCT and hsCRP in prediction of blood stream bacteraemia in patients presenting to ED.
General comments:
1.	Patients included in this study were treated in 2010 onward, specify the end date.
2.	The language used across the manuscript, particularly in the abstract, need significant English editing.
3.	Sentences such as “hsCRP should be abandoned ……” should be avoided and deleted.
4.	The short title should reflect the biomarker interest. Suggest changing to Biomarkers of sepsis in adult emergency.
5.	Why there is no ED physicians included as authors?
6.	A sensitivity analysis should have been performed on patients who had one or 2 tests done not just those who have had ALL the 3 tests done.
Specific comments:
1.	The mini VIDAS biological range is much lower than 0.5 ng/ml. I thought it goes down to
0.06.

Methods: It is not helpful having too many categories of culture negative. Stick with a definition and just use the total number of what was categorised as negative culture in the analysis.

In the results, graphs and tables.
1.	 It is very adequate to ONLY separate g-ve and g+ve. No need for ROC for bacilli and cocci. This should apply to all results including the tables. There is little pint dividing cultures into 4 groups in addition to negative, contaminant and others.
2.	PCT cut off was chosen at 0.5 ng/ml. Do you have data for PCT cut off at 0.25 or 0.1 ng/ml? In addition is there an upper cut off value for best performance of PCT?
3.	Suggest merging tables 1, 2 an 3 into one table.
4.	Table 7 is not necessary. This is not a MA. I suggest comparing the results with your data in the discussion section.
Discussion should be restructured to address the following:
1.	Start with key results
2.	Explain why DOR of combinations is better than single DOR.
3.	Compare with previous trials
4.	Implication of the above finding for ED practice and sepsis management in general
5.	Limitations of the study
6.	Next step.
7.	Shorter conclusion (should not have any results in the conclusion).


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: DIAGNOSTIC VALUE OF SERUM PROCALCITONIN, LACTATE, AND HIGH-SENSITIVITY C-REACTIVE PROTEIN FOR PREDICTING BACTEREMIA IN ADULT PATIENTS IN THE EMERGENCY DEPARTMENT
Review round: 2
Reviewer: 1

Output:
Basic reporting: It is still a lack of literature references and context in the Discussion. Only a very few of your results are compared with other peoples work (and here only two other works are cited). The Disucssion must be extended and the obtained results put in a context, i.e., compared with previous studies reporting the performance of the single biomarkers and/or combined biomarkers for predicting positive blood cultures. Reasons for differences/similiarities in findings between studies?

Line 249 and 257: No information about the frequency of positive blood cultures for the expanded cohorts. This is important as the group sizes are very important for the performance characteristics. This information must be added in Method and Material section.

Line 184-186: this sentence is very hard to understand and must be rewritten.

Line 197- : all the data presented in the text here should be presented by tables only.

Line 246: threshold for PCT is not defined here.

Inconsistent use of hs-CRP vs. high-sensitive CRP. This should be looked over throughout the manuscript.
Experimental design: No comments.
Validity of the findings: In the Abstract: You write "Among patients with elevated PCT and/or LAC, the odds for predicting positive blood culture was nearly five-fold higher compared to that in patients with normal levels (diagnostic odds ratio, 4.83; 95% CI, 42 2.84–8.69). The diagnostic odds ratio was nearly eight-fold higher for the prediction of positive gram-negative bacteria (GNB) culture (diagnostic odds ratio, 7.75; 95% CI, 3.56–20.03) and it was nearly three-fold higher for the prediction of positive gram-positive bacteria (GPB) culture (diagnostic odds ratio, 2.87; 95% CI, 1.43–6.41)." I wonder where these figures come from as I don´t recognize the from the results reported in the manuscript? The abstract should only report exactly the same results as in the Results section.

A diagnostic accuracy of only 65% is not acceptable from a clinical point of view (as stated for example in line 275 and line 347). This is considered to be rather poor actually.

It is wrong to say that CRP should be abandoned from the ED; it is still useful as a marker for bacterial infections but is limited by it´s slow kinetics and should be viewed from the timpe point of disease onset.
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: DIAGNOSTIC VALUE OF SERUM PROCALCITONIN, LACTATE, AND HIGH-SENSITIVITY C-REACTIVE PROTEIN FOR PREDICTING BACTEREMIA IN ADULT PATIENTS IN THE EMERGENCY DEPARTMENT
Review round: 2
Reviewer: 2

Output:
Basic reporting: Nil
Experimental design: Nil
Validity of the findings: Nil
Additional comments: Thank you for a very improved manuscript.
I have one suggestion to remove the number of patients from the title.