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: DE NOVO OVIDUCT TRANSCRIPTOME OF THE MOOR FROG RANA ARVALIS: A QUEST FOR MATERNAL EFFECT CANDIDATE GENES
Review round: 1
Reviewer: 3

Output:
Basic reporting: Apart from a couple of typos (which I have included in the "Comments for the author" section) the manuscript is written in a clear, concise, self-contained and professional manner. Literature is well cited, data is well presented in figures.
Experimental design: This is a good first step in the process of identifying the desired genes. But I think a comparison of these oviduct-specific transcriptomes with transcriptomes from other tissues is needed in the future to find oviduct-specific genes or expression profiles. That said, this paper still represents a valuable resource.
Validity of the findings: no comment
Additional comments: Overall it is a solid paper - descriptive (necessarily so), but well executed and a useful resource.

Some more specific comments relating to the writing of the manuscript:

>>Introduction

104 - consider “encompassing” instead of “bracketing”  here

108 - first set of transcriptomic markers, maybe, but not the first set of genome-wide markers. See this paper, where they generate RADseq markers across the genome:

Brelsford, A., Lavanchy, G., Sermier, R., Rausch, A. & Perrin, N. Identifying homomorphic sex chromosomes from wild-caught adults with limited genomic resources. Mol. Ecol. Resour. 17, 752–759 (2016).

Please amend the sentence to reflect this. Thanks.

>>Methods

185 - Kyoto Encyclopedia of Genes and Genomes (KEGG). You use KEGG earlier (167) - add the acronym definition there instead.

>> Results

No issues here

>> Discussion

320 - I think you mean liver fluke here.

319 - 324 - your suggestions for the fluke hits are as follows: i) due to poor annotation of amphibian genomes (I presume you are implying that the genes were therefore missed by the annotations of their genomes), ii) orthologous with genes of the liver fluke, or iii) R. arvalis may have been infested by other genetically related parasites.

I don’t quite agree with the priority given to these hypotheses, see below.

Re i): Given the quality of the Xenopus genomes, I would say that it is unlikely that so many genes would be missed by their annotation.
Re ii): The argument that these genes are orthologous to Platyhelminthes would also require that they are absent in the other existing amphibian (or any vertebrate) genomes for these to be the best hits for those genes. So that, to me is almost the same as i).
Re iii): For me this is the most likely explanation.

So, perhaps you could even just leave i) and ii) out, if the genes were absent from other amphibian genomes, I would not expect the next best hits to be Platyhelminthes, so I don’t think this result has occurred by accident. A useful thing to look at would be how much of the fluke genome these unigenes represent. It might be that you have actually produced two transcriptomes here, which would be cool.

384 - “lay” not “lays”


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: OBESITY, BUT NOT HYPOHYDRATION, MEDIATES CHANGES IN MENTAL TASK LOAD DURING PASSIVE HEATING IN FEMALES
Review round: 1
Reviewer: 1

Output:
Basic reporting: Tables 2, 3 and 4 don`t present p values and they should, since they intend to demonstrate  the differences between analyzed groups and situations. The figures need explanatory footnote, however, I would consider deleting them, because they repeat data already described both in the tables and the text.
Experimental design: No comment.
Validity of the findings: No comment.
Additional comments: There are some phrases along the manuscript that lack the initial capital letter (lines 52 and 351). Some terms appear unabbreviated again, after their acronyms have already been explained (lines 259 and 260).  In the discussion (line 353) Kempton et al suggest (not "suggests" since et al means and others). In the conclusion (line 433) the word "to" is missing...effort TO maintain.


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: OBESITY, BUT NOT HYPOHYDRATION, MEDIATES CHANGES IN MENTAL TASK LOAD DURING PASSIVE HEATING IN FEMALES
Review round: 1
Reviewer: 2

Output:
Basic reporting: No comment.
Experimental design: No comments.
The experimental design is clear and seems appropriate to respond the objective of the present study. The main question is why authors choose hypohydration equal to 2 percent normal body mass and hyphertemia equal 1o C to compare differences in cognition, mental task load and mood between obese and non-obese female. What physiological or perceptual response could impair or improve these variables differently, based on a heterogeneous group in body composition. This may not be a weakness of this study, but it might be included in the rationale.
Validity of the findings: No comments.
Additional comments: Manuscript entitled “Obesity, but not hypohydration, mediates changes in mental task load during passive heating in females” (#25543)

General comments

This manuscript makes an important contribution to the literature in the area by providing information regarding the effects of hypohydration and hyperthermia on cognition, and mood in obese and non-obese. The objective of this manuscript, as described by authors, was to examine the independent and combined effects of mild hypohydration (i.e., <2% body mass loss) and hyperthermia on cognition, mental task load, and mood in obese and non-obese individuals. My suggestion is that the introduction and discussion must be reviewed to include physiological mechanisms hypothesis for the results found in the present study, mainly related to obesity.

Comments to the Author

1)	Hydration: The authors chose to examine the impact of an approximate dehydration level equal to 2 percent normal body mass.  A number of manuscripts were cited, but most did not indicate which % of hypohydration showed or not impairment on cognitive performance or impact on mental task load or  mood. For example, Szinnai, Schachinger et al. 2005 showed cognitive-motor function is preserved during water deprivation in young humans up to a moderate dehydration level of 2.6% of body weight. Based on available literature, what basis did they have to suspect a difference in cognition, mental task load and mood at 2% body mass loss?

2)	 Population: In addition, it was not clearly explained the possible physiological differences in obese population that could affect these variables in comparison on non-obese individuals during or after water deprivation and/or heat stress. Why study this population regarding these aspects? One explanation brought is that military personnel, construction workers, and firefighters, are often exposed to occupational heat stress. Also, about one-third of Americans are reported to be overweight or obese. Please, clarify these point.

3)	Hyperthermia: Possible explanation for thermoregulatory differences is that obese have a smaller BSA per unit of body mass than non-obese, which could affect their ability to effectively exchange heat with the environment. Since fat has a specific heat that is approximately half of that of fat-free mass, obese also can experience an insulative effect where a given heat load will increase their body temperature more than that of a lean individual (Buskirk et al. 1969). These possible differences between obese and non-obese are controlled in the present study as hydration levels and hyperthermia were the same between groups. Which physiological other physiological differences could affect cognitive responses, on mental task load and mood between groups? Please, include in the manuscript.

Specific Comments

4) Introduction. Line 73. However, other studies report no cognitive differences with mild hypohydration (~2%) when fluid is restricted for 36-h (Szinnai, Schachinger et al. 2005). Please, correct this sentence (one study or include other references).
        In addition, how can this design lead to more hydration specific recommendations, when literature available suggests few or no effect will be detected at 2% body mass loss?

5) Methods. Line 101. Please, include additional information about sample size calculation.

6) Discussion. Line 325. What is the possible explanation for that a similar increase in body temperature resulted in an increased mental task in obese, but not non-obese?

7) The conclusion could be more of a take away message rather than an abrupt end more reflecting a result as it currently reads.


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: OBESITY, BUT NOT HYPOHYDRATION, MEDIATES CHANGES IN MENTAL TASK LOAD DURING PASSIVE HEATING IN FEMALES
Review round: 1
Reviewer: 3

Output:
Basic reporting: Please see comment in section 2 about references pertaining to head/brain heating and cognitive function.
Experimental design: 1.	N=21 (10 obese, 11 non); What kind of power did the authors have?  Please provide this.
2.    Does serum osmality/urine osmolality or Usg vary considerably during follicular phase?please comment.
3.	“Learning effect” with ImPACT?  (48 hr b/t trials) 5 parallel formats?
4.	Replicated diet portion (calories) for subsequent trial… macro/micronutrient content could impact hydration state…?  No mention was made by authors to address nutrient composition stated replication of diet (via caloric content)
5.	“refrain from high moisture content foods” 24 hr prior in dehydration; was there significant difference prior to start of two trials?  Why no baseline measures for Table 2, would they hold value to prove hydration levels against trial and not just pre/post a “control” so to speak ? Table 2 does not show statistical significance… were there no differences?
6.	The water suit…  head was not necessarily at same temp, could sweat while suit precludes sweat from body...this may be critical for cognitive study.  What was the temperature of the brain?  I know you did not measure this, but i needs to be addressed as your cognitive assessment in measuring brain function and we have no references to what this intervention does to the head/hippocampus/brain.  This should be acknowledged as a limitation (also given the fact that the “global application” in the intro is around firefighters/military/athletes in their examples do not have their heads excluded from exposure).
7.	Mood assessment after instrumentation/rectal probe, in suit? None taken on baseline day to determine if suit/positioning alone has impact on “mood” considering if instrumentation is a factor.
Validity of the findings: 1.	Statistical Analysis (Line 222):  series of separate univariate ANOVA’s performed for all physiological (lists), hydration (lists).  Reviewer notes this sentence is incomplete, believe word “variables” or similar may be missing.

2.  What dose of heat increase occurred in brain?  Please add any references if possible
Additional comments: 1.	Line 351 Capitalize Y in young


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: OBESITY, BUT NOT HYPOHYDRATION, MEDIATES CHANGES IN MENTAL TASK LOAD DURING PASSIVE HEATING IN FEMALES
Review round: 2
Reviewer: 1

Output:
Basic reporting: No comment.
Experimental design: No comment.
Validity of the findings: No comment.
Additional comments: The requested changes have been made. No additional comments.


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

Input:
Title: SPATIAL AND TEMPORAL SHIFTS IN THE DIET OF THE BARNACLE AMPHIBALANUS EBURNEUS WITHIN A SUBTROPICAL ESTUARY
Review round: 1
Reviewer: 1

Output:
Basic reporting: The authors present a clearly written, well-thought out manuscript on the generalist feeding strategy of barnacles across a broad spatial scale over the course of a year.  Overall, the manuscript reads very well, although there are a few spots which I think could receive a little more attention.

LN 87-90: Can you provide a range (if available) of particle sizes that barnacles can effectively remove from the water column?
Ln 97-99: I think there is a word missing in this sentence before location?
LN 103-107: What was particularly devastating about these blooms? Are they toxic? What is the size of the brown tide cells? It would be beneficial to provide a little more details on the bloom species.
LN 238: Does decoupling occur during the brown tide blooms? It looks like it might.
LN 242: I am curious about the C signature in the tissues here (and I have also brought this up below) - is there any literature on barnacle tissue turnover time? isotopic signatures are integrated over time, and while the N may stay more stable if the sources of N are chronic, I wonder if the variability in the C isotopes has anything to do with tissue turnover/growth rates. Thoughts?
LN 288-290: Is this enrichment also possibly indicative of a second trophic step, perhaps that during bloom conditions, the barnacles do feed on zooplankton? Is it possible that despite the generalist feeding nature of barnacles that the brown tide is too small to be captured by their filter?
Experimental design: The authors use a sound research approach, particularly as it pertains to hypothesis 1 - that isotopic values vary over space and time and will be closely linked to POM.  I am less convinced of the approach to hypothesis 2 in effectively showing that the barnacles are consuming the brown tide plankton.  I don't think this is a major issue, just one that should be discussed further - based on the isotope data presented alone, it is not clear that the barnacles consume the brown tide. Experiments would have been more useful in showing this.
Additionally, a little more detail on the analysis - particularly, it is not clear if bloom samples were included in the regression analysis (also unclear if they should be), or whether the bloom samples were analyzed separately (which would at least add more teeth to the argument about barnacles consuming brown tide).  A little clarification here would be beneficial.
Validity of the findings: As previously mentioned, this was a sound research approach, and particularly the temporal and spatial exploration of barnacle tissue and POM isotopic values is robust and technically sound.  The authors were able to demonstrate high spatial and temporal variability in isotopic signatures of POM and barnacle tissues, and despite this variability, the two tracked each other very well, especially during non-bloom periods. This was especially true for nitrogen, but less so for carbon. I have a few questions regarding this - there is considerably more variability in the C signatures. Do the authors have any ideas as to why? This could receive a little more attention in the discussion.  Additionally, is there any information about the tissue turnover times of barnacles in general, or this species in particular? The authors present compelling evidence regarding the fairly steady N conditions in the IRL, but the C source likely changes quite a bit over time, and the isotopic signature of tissues is reflective of not only the source of C, but the source integrated over some period of time, and so there might be a lag in the C signature of the POM and the tissues.  Can this be expanded upon?

The effects during the bloom are a little less convincing - yes there is changes in the isotopic signatures of tissues compared to the annual mean, but without specific analysis of this, I am not sure the conclusions are very well supported by the data.  I think there is an interesting story here - and there does appear to be a shift in signature during these bloom periods, but I am not sure if this is because they are consuming the brown tide or if there is some selectivity during this time period.  Could the authors do analysis of just the brown tide samples similar to the other POM samples to show that the tissue still tracks the POM during the brown tide? Further, the N signatures during the brown tide are enriched 6-7 permille, which might be more indicative of a 2 step trophic shift.  This should at least be discussed as a possibility.
Additional comments: I found this manuscript to be well-written and thorough.  I feel the issues I have presented here should be very easy to address in a minor revision. I commend the authors on their good 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: SPATIAL AND TEMPORAL SHIFTS IN THE DIET OF THE BARNACLE AMPHIBALANUS EBURNEUS WITHIN A SUBTROPICAL ESTUARY
Review round: 1
Reviewer: 2

Output:
Basic reporting: This manuscript is very well written, and the authors represent previous literature well within their Introduction. The article is structured appropriately and the raw data is shared. The hypotheses are clearly stated and the results are relevant to the hypotheses.
Experimental design: The study at hand is a sound investigation of barnacle diet, focusing on the seasonal variability in these diets. The researchers’ additional interest in anthropogenic effects, in the form of algal blooms, further contributes to the scientific understanding of human impacts on coastal species and the ability of those species to adapt to perturbations.

The study question and hypotheses are well defined by the authors. However, the justification provided for the question is slightly mismatched from the study design. To truly address the study goal [(ll. 109-112) “To determine whether the abundance of A. eburneus across sites in the NIRL is linked to generalist feeding on available sources of suspended organic matter…”], the researchers should have not only assessed the diets of Amphibalanus eburneus, but also the diets of the other less dominant suspension feeders in the community. This can be remedied by simply rewording the study goal.

Both field collections and laboratory procedures were conducted appropriately and with the necessary permits. Methods are described in sufficient detail for replication.
Validity of the findings: The seasonal representation of POM values in IRL is a valuable contribution to the ecological understanding of the area. It also provides information about the level of human impact on IRL sites.

The evaluated coupling of POM and A. eburneus tissue values is warranted and conclusions are valid given the reported results.

The authors’ conclusions breakdown a bit when they begin to justify zooplankton as a lesser component of A. eburneus diets. Though I agree that the authors would have seen an additional trophic step between POM and A. eburneus tissue if the barnacles were consuming large quantities of zooplankton, I did not follow the authors’ logic about body size (ll. 258-261). If the authors are positing that larger body size correlates with feeding at a higher trophic level in barnacles, references to previous literature showing as much needs to be included. The following line of evidence noted – the strong correlation between 15N of A. eburneus and salinity – is similarly diffuse and requires more explanation and reference to the literature (you do this well in the Abstract ll. 32-34). The statements presented in ll. 264-273 present much clearer evidence for the validity of the authors’ findings.

The Conclusion contains statements that link the findings of the present study to a conclusion that, “The generalist feeding strategy of A. eburneus...likely contributes to the successful dominance of this species across sites in the NIR,” (ll. 300-302). Though this may be true, I do not think the authors can make this claim as a central conclusion given that their methods did fully address the question backing this statement (see my comments in Experimental Design). This point can still be made, but it should not be considered “likely”.
Additional comments: In addition to addressing the points raised in the previous sections, please address the edits listed below:

ll. 107-109 and Figure S1 – Why are amphipods included in this? Are they tube dwelling amphipods that create substrate? Additionally, the categories are rather different between 2014 and 2016 – were the same sites used? It’s ok if they’re not, but the authors should state this clearly. Finally, you say “suspension feeders”, but you have included algae – please justify.
l. 130 – Start a new paragraph with “To test whether a bloom…”
l. 346 – Add a comma after “Deegan”.
ll. 359-363 – Switch the position of these two citations.
l. 392 – Add a period after the pages.
ll. 393-394 – Title of the article should not be in caps for all words.
l. 411 – Add comma after MacKenzie.
l. 412 – “eds.” Needs to be consistently capitalized or lower case throughout citations.
l. 419 – Add comma after Michener.
l. 426 – Add comma after Phlips.
l. 429 – Remove comma after Coasts.
l. 430 – You previously capitalized the word after the semicolon in l. 428. Keep it consistent.
l. 431 – Remove comma after Ecology.
l. 440 – Keep capitalization after semicolons consistent throughout citations.

Figure 5 caption – Add period after “clarity”.
Figure S4 – I think the manuscript would benefit from this figure being moved to the main text. This gives the reader a much more visually succinct understanding of how A. eburneus tissue values change with season and algal bloom.


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: TESTING ‘PROPORTION OF FEMALES CALVING’ AS AN INDICATOR FOR POPULATION-LEVEL REPRODUCTIVE PERFORMANCE FOR BLACK RHINOCEROS (DICEROS BICORNIS)
Review round: 1
Reviewer: 1

Output:
Basic reporting: 1.	The manuscript was generally well-written using professional English. I believe, however, there was an excessive use of non-standard acronyms (e.g., PFC, IBI, AFR, ACF) that made for difficult reading, and required flipping back and forth to find the meaning of particular acronyms.
2.	The manuscript provided a reasonable background with appropriate literature citations. I was surprised that the authors didn’t use the IUCN status of black rhinoceros (Critically Endangered, but with increasing populations) as a justification for their research.
3.	The manuscript conformed to PeerJ standards.
4.	The figures were clear, but I didn’t see the need to provide the transformations along with the original data in Fig. 1, which cluttered the figure. Also the Y-axis on Fig. 1 should be labeled. I believe it would be helpful to present the Percent of Females Calving in conjunction with yearly data on population size in the same figure (data on population size are only available in the supplemental data). The time-series analysis in Fig. 2 was inventive, but the levels of significance represented by the horizontal dashed lines needs to be identified.
5.	Raw data were provided in a table.
Experimental design: 1.	This original research falls within the scope of the Journal. The authors are to be commended for their long data run, which is unusual for large mammals.
2.	The research question was relevant; there are few data on asynchronous non-seasonable breeders with an inter-birth interval of more than 1 year. A major shortcoming, however, was the failure to set forth a set of hypotheses at the end of the Introduction and the criteria necessary to test them. Framing the Discussion around those hypotheses also would improve the organization of that section, and more clearly indicate how the knowledge gap would be filled by this study.
3.	The research was rigorous and likely performed to high ethical standards, although I would like to have seen an animal care and use approval (or necessary permits) for the original research, even though the authors were just analyzing data in this paper. The Methods were clear, but the authors relied heavily on previous publications, and providing a bit more information would have been useful.
Validity of the findings: 1.	 There are several ways to use AIC modeling. I wonder what the analysis would have looked like if only the top 3 models (those with changes in AIC of less than 2 from the top model) were averaged. The influence of precipitation is discounted, but rain occurs in one of those top models.
2.	Some evidence of density dependence was reported in their previous publications—a relationship occurred between age at first reproduction and population density. The critical point is not density per se, but density in relation to ecological carrying capacity (K). Parametrizing where the population is with respect to K is necessary to see how percent of females calving might change as the population approaches K. I doubt there are data in this manuscript to allow that assessment, but the Discussion would be improved by a more complete discussion of ungulate population dynamics and some predictions about how things might change in the future as density continues to increase.
Additional comments: 1.	Long data runs of this type are unique and critical to our understanding of population dynamics and metrics that reflect the status of populations.
2.	A clear-cut approach to stating and testing hypotheses would provide a more coherent paper with less ambiguous conclusions.
3.	Placing this research in a firmer context of ungulate population dynamics would improve the scope of the paper and help with identifying gaps in our knowledge.


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: TESTING ‘PROPORTION OF FEMALES CALVING’ AS AN INDICATOR FOR POPULATION-LEVEL REPRODUCTIVE PERFORMANCE FOR BLACK RHINOCEROS (DICEROS BICORNIS)
Review round: 1
Reviewer: 2

Output:
Basic reporting: This manuscript addresses a specific aspect of population monitoring using a detailed and impressive data set from a reintroduced population of black rhinos.  As such, the work is likely useful for managers of populations in rhinos.

The language is generally clear and concise, however, there are several aspects of the organization and writing that would benefit from revision.
1.	The title does not provide a reader with a clear understanding of the content of the paper without reading at least the Abstract because PFC is not a common abbreviation.
2.	Similarly, although the population name (SKKR) is “not an acronym of an official name”, it is distracting, leaving the reader to wonder what it does stand for.
3.	Figure and Table legends and captions are too vague/brief to allow readers to understand the data presented and its significance.  More context would help with interpretation of the information presented.  For example, in Table 1 the abbreviations should be identified either in the caption or in a footnote.
4.	The Introduction could do a better job of setting up the problem and providing context for why this topic should be of interest to a broad readership.  The topic is so narrow that it comes across almost as a rebuttal than a presentation of research results in a stand along article.  This is apparent when discussing benchmarks for populations (Lines 50-54 and again in the Discussion in the paragraph beginning with line 176).
5.	A large number of statements/conclusions are presented in the Introduction without data or citations for where the data could be found (e.g., statements in lines 63-71 and 81-83).  This also contributes to the feeling that this short paper is an addendum rather than a complete study or analysis.

A few specific comments/suggestions for improvement:
Line 128:  This paragraph could use some context and a strong topic sentence.
Line 134:  This statement and citation are more appropriate for Discussion than Results.
Line 137-138:  This information and citation belong in the Methods.  The AIC weights would be better presented in table.
Line 144-146:  What criteria was used to make this decision?
Experimental design: Data on population size, sex ratios, reintroduction history, and stage structure over the study period would help with interpretation of the analyses.  The reliance on previously published papers for this information requires that readers trust the statements without an opportunity to evaluate them.
Validity of the findings: The data and analyses are relatively simple, however, there are a couple of points regarding interpretation that could use additional thought.

1.	The sample size of adult females is quite small during the study period (<8 individuals for the first half).  The high stochastic variation in reproduction during the early time period should not be surprising given the small number of individuals, and the reduction in variation overtime likely reflects, in part, an increase in sample size.  This temporal trend should be discussed/acknowledged in the Discussion, and it might be more transparent to include the numbers of adult females in the text or perhaps in the figure legend (that information is included in the supplementary table).
2.	A related comment regarding interpretation is that the long intervals between births (~2.5 yr) should result in the lag effects in reproduction given even a modest amount of synchrony among females.  It almost would be surprising if this was not the case.
3.	Density is not important in this population yet because it is not close to carrying capacity.  How does this influence interpretation of your results?
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: TESTING ‘PROPORTION OF FEMALES CALVING’ AS AN INDICATOR FOR POPULATION-LEVEL REPRODUCTIVE PERFORMANCE FOR BLACK RHINOCEROS (DICEROS BICORNIS)
Review round: 2
Reviewer: 1

Output:
Basic reporting: The hypothesis testing was improved.
Experimental design: The authors satisfied my concerns with their analysis.
Validity of the findings: I thought the authors did a better job of casting findings from their research in a broader framework that acknowledged the potential value of their findings.
Additional comments: The authors have done a reasonable job of revising their manuscript, and satisfied most of my concerns.

There remain a few small corrections:
Lines 100-105—break this long sentence into several clear sentences.
Lines 110, 139—data are plural and these should read “data are” not data is.

There is one point, however, that I would like to see resolved. The authors point out that data used in their manuscript were collected as a result of management objectives rather than a research project. I do not believe that his absolves them, however, from indicating that data were collected in a responsible manner. The manuscript is research and it is their responsibility to indicate whether data they used were collected in a conscientious fashion reflecting the ethical treatment of animals. Perhaps a way to accomplish this would be to indicate that data were collected in keeping with guidelines for research on wild mammals (Sikes et al. 2011, Journal of Mammalogy 92:235-253).


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: TESTING ‘PROPORTION OF FEMALES CALVING’ AS AN INDICATOR FOR POPULATION-LEVEL REPRODUCTIVE PERFORMANCE FOR BLACK RHINOCEROS (DICEROS BICORNIS)
Review round: 2
Reviewer: 2

Output:
Basic reporting: Revision addresses comments.
Experimental design: Revision addresses comments.
Validity of the findings: Revision addresses comments.
Additional comments: Although the rebuttal has a somewhat argumentative approach, the authors have addressed my concerns and comments sufficiently.


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: THE APPROPRIATENESS OF CEFTRIAXONE AND METRONIDAZOLE AS EMPIRICAL THERAPY IN MANAGING COMPLICATED INTRA-ABDOMINAL INFECTION—EXPERIENCE FROM WESTERN HEALTH, AUSTRALIA
Review round: 1
Reviewer: 1

Output:
Basic reporting: This retrospective study has a clear aim - to identify the pathogens involved in common surgical conditions that lead to intra-abdominal abscesses. They also nicely showed whether their recommended antibiotic use was appropriate treatment for those same organisms due to their sensitivities.

The article is generally clear and unambiguous; there are a few grammatical errors which require addressing and have been listed below.

The Introduction is concise and sets the scene well. More recent literature references could be added (please see below).

The references are generally up to date and appropriate

The figures are good and are both relevant and well labelled.
Experimental design: The authors have nicely presented a single centre experience in the microorganism presentation for surgical intra abdominal abscesses. There are some larger studies in the literature but they tend to only cover specific conditions rather than the whole spectrum which the authors have done.

The research question is well defined and the results are simple but clear and relevant to common practice.

There are weaknesses in this study; it is reliant on DRG coding and when they subdivided the conditions causing intra abdominal abscesses, some of the groups were of smaller numbers (eg cholecystitis group n= 9)
Validity of the findings: Although this is a retrospective study, the search they performed is sound and they have produced a descriptive analysis of their results.

The data is robust.

The conclusion is supported by the results which they identified is correct - they have illustrated that in more complex perforations, there is a higher likelihood of the presence of resistant organisms and therefore require consideration of an alternative antibiotic regime.
Additional comments: The strengths of this study are that it provides a summary of organisms and their sensitivities in a range of surgical conditions from a single treating institution. I think that the take home message is clear and is translatable to clinical practice.

I would support this articles acceptance, but with the following conditions are addressed / answered:-

1.	Patients who had resistant organisms - were there any underlying risk factors for these (previous recurrent admissions for antibiotic therapy for the same condition)
2.	Did all patients who were sensitive to Ceftriaxone and Metronidazole respond to this therapy or were there patients who had to have second line antibiotic therapy

3.	Line 73 - there is more recent literature evidence supporting that the delay of sepsis control is associated with poorer outcomes.
4.	Line 84 - grammatical change - In this context two clinical questions arise ;- 1. Whether empirical therapy using ceftriaxone and metronidazole provides sufficient antimicrobial coverage in IAI overall and 2………
5.	Line 209 - to correct sentence IAI - this includes the "use of" ceftriaxone

6.	Line 224 - What is the definition of "higher risk patients"…


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

Input:
Title: THE APPROPRIATENESS OF CEFTRIAXONE AND METRONIDAZOLE AS EMPIRICAL THERAPY IN MANAGING COMPLICATED INTRA-ABDOMINAL INFECTION—EXPERIENCE FROM WESTERN HEALTH, AUSTRALIA
Review round: 1
Reviewer: 2

Output:
Basic reporting: the authors don't stress between un complicated and complicated IAI
and the relative involving bacteria and manage and microbiological sampling
the epidemiological and/or local incidence of ESBL and MBL bacteria in the described setting is not showed
Experimental design: the aim and scope should be include the  difference above cited: describe the epidemiology of setting, the kind of infections complicated and uncomplicated
characteristics of population comorbidities, previous hospitalization ect
Validity of the findings: the manuscript showed the efforts of authors to find an empirical treatment of cip antibiotic treatment despite the field is very difficult to manage
Additional comments: I appreciate the efforts of the authoris but the matter it is so complex that deserves to characterize better the surgical aspect, intervention in emergency or election, local epidemiology and to comment with help of the literature if this empirical therapy can be applied only in your hospital or is an option for setting with the same epidemiological microbiological isolates


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: THE APPROPRIATENESS OF CEFTRIAXONE AND METRONIDAZOLE AS EMPIRICAL THERAPY IN MANAGING COMPLICATED INTRA-ABDOMINAL INFECTION—EXPERIENCE FROM WESTERN HEALTH, AUSTRALIA
Review round: 2
Reviewer: 1

Output:
Basic reporting: .
Experimental design: .
Validity of the findings: .
Additional comments: As per my previous review - I was generally happy and feel a further review not necessary on my side.


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: THE EXPRESSION AND CLINICAL SIGNIFICANCE OF GADD45A IN BREAST CANCER PATIENTS
Review round: 1
Reviewer: 1

Output:
Basic reporting: The manuscript  “The expression and clinical significance of GADD45A in triple negative breast cancer” by Wang et al. presents the expression of GADD45A in breast cancer as potential new prognostic biomarker in particular in triple negative subtype. Breast Cancer remains the most common malignancy and the major cause of cancer death among women worldwide and new achievements in prevention and treatment, including reliable new prognostic and predictive biomarkers, are still required. This study is therefore well focused on the current clinical needs.

However some issues had to be addressed for publication.

The first concerns regards the English language used, that should be improved to ensure a can clear understanding of the text and the conclusions. Some paragraphs (such as lines 172-181) are confused and re-phrasing could be useful to clearly explain and correlate the results with previous findings.  I suggest a complete review of manuscript aimed to better organize the sentences and  support the conclusions. Some editings are needed as well (grammar (ex line 198) or table (Ki67 value in Table 4).

The literature listed is sufficient to describe the state of the art and define the study goals, but a deeper discussion of the current results in comparison with previous finding is needed. In particular, authors seems to claim that this is the first study where clinicopathologic features are correlated with GADD45A expression. However, as they correctly reported, Tront et al. in 2013 performed similar investigations. For example both the studies demonstrated that GADD45A expression is higher in tumor  compared to normal/adjacent tissue. I suggest to insert some sentences to put in evidence the differences between the two studies and to underline the novelty of their own.
The paper results are consistent with the study goals.
Raw data are provided as for PeerJ policy requirements (even only 1 TMA is shown); ethical documentation presented only in Chinese.
Experimental design: The study aims to correlate GADD45A expression obtained by IHC analysis to clinical pathological feature. This a basic research that potentially could add new information for the understanding of breast cancer biology. It has the great advantage to present a large cases collection, well divided in subtypes, with a long patients’ follow up.
Material and Methods:
Correct parameters and references are reported for breast cancer characterization (the definition/evaluation of ER/PgR and Her 2 expression/positivity are the currently used in clinic).
However I have some concerns about the Ki67 cut off. Authors classify Ki67 index> 14% as high. The accepted cut off value for ki67 is 20%, as from St Gallen consensus (Personalizing the treatment of women with early breast cancer: highlights of the St Gallen International Expert Consensus on the Primary Therapy of Early Breast Cancer 2013. Goldhirsch A, Winer EP, Coates AS, Gelber RD, Piccart-Gebhart M, Thürlimann B, Senn HJ; Panel members. Ann Oncol. 2013 Sep;24(9):2206-23.)
Therefore this value should be used to discriminate between high and low Ki67 expression. Could the authors explain the reason why they choose 14% as Ki67 cut off? This value is quite important since from their analysys this parameter correlates with GADD45A expression. On note, strangely almost half of triple negative cases analysed in this work (30 vs a total of 79 tissues) present ki67 values lower that 14%, while, as the authors remarked in the discussion, ki67 is normally high in triple negative breast cancer.
Regarding ICH analysis, material and methods section is quite vague. A very poor description of the procedure is presented (line 839 “standard protocol” should be better described).
Moreover normally sections are analysed in triplicate by at least two independent pathologists. It’s not evident if the authors follow this correct laboratory practice to analyse the tissues.
Validity of the findings: The authors conclude that ER negative, high histological grade and high Ki-67 index correlate with high GADD45A expression. These conclusions are opposite to the ones presented by Tront et al (2013) who showed that high GADD45A levels are associated with luminal A and luminal B subtypes and that in triple negative tissue this expression is low. In the same way Tront and coworkers didn’t find any correlation with the histological grade, that here resulted significant.
Could the authors provide an explanation for these differences?
This discrepancy could be generated by the higher number of tissues analyzed in this work compared to the previous one (419 vs 45). Wang et al were also able to shown the correlation with PgR and ER separately while Tront et al analysed only tissue where they were both positive. This point could be taken into account to discuss the results. Comments on these data could be inserted in the manuscript.
To add novel information to the field I suggest these majors
-	Provide correlation  with breast cancer subtypes. The information about GADD45A expression in regards with the different subtypes could be important and useful for clinicians.
-	Repeat the statistical analysis considering ki67 cut off > 20% to confirm the data.
-	Include a multivariate analysis. The author stated that GADD45A is an independent prognostic value for triple negative breast cancer subtypes. However, as they put in evidence in the discussion (line 184), this could be an effect of ki67 value. The author performed only univariate Pearson χ2 test or Fisher´s exact test to confirm the correlations between clinicopathological factors and GADD45A expression. A multivariate analysis is recommended to strengthen and confirm that GADD45A could be considered a prognostic marker for triple negative breast cancer.
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: THE EXPRESSION AND CLINICAL SIGNIFICANCE OF GADD45A IN BREAST CANCER PATIENTS
Review round: 1
Reviewer: 2

Output:
Basic reporting: No comment
Experimental design: I suggest to report if the series are consecutive
Validity of the findings: No comment
Additional comments: The paper is interesting but I suggest to mitigate the emphasis on the prognostic role in TNBC because of the small sample size


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: THE EXPRESSION AND CLINICAL SIGNIFICANCE OF GADD45A IN BREAST CANCER PATIENTS
Review round: 1
Reviewer: 3

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: This manuscript identified GADD45A as a potential treatment target by performing immunohistochemistry in 419 breast cancer tissues and 116 adjacent non-neoplastic tissues. Although this manuscript has assessed a number of patient samples, this review has concerns of their less developed experiments and less reasonable conclusions.
GADD45 is known as a tumor suppressor. Although it is induced in breast cancer, it does not conclude that GADD45A is a druggable target of cancer treatment. The function of GADD45A should be addressed with further experiments. In addition, it is also important to check the mutations in breast cancer. If GADD45A is frequently mutated or GADD45A has new oncogenic functions in breast cancer especially TNBC, the assumption of the authors may be possible.
The authors refer the publication of GADD45A-promoted MYC-driven cancer to explain their result and hypothesis. However, GADD45A is more well-known as a negatively regulated MYC downstream target.
Furthermore, the clinical significance of GADD45A in TNBC must be intensively validated to overcome all allegations of GADD45A role in breast cancer.
Overall, this review concludes that this manuscript is not sufficiently developed and described to be published in PeerJ.


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: THE EXPRESSION AND CLINICAL SIGNIFICANCE OF GADD45A IN BREAST CANCER PATIENTS
Review round: 2
Reviewer: 1

Output:
Basic reporting: Wang and co-workers evaluated the expression of GADD45A in breast cancer using IHC analysis.

One of the main concerns regarding the manuscript was the English language used, since some paragraphs were poorly written and confused. The authors deeply revised the text improving the language, making more understandable data description and relative conclusions. Some corrections are still necessary (ex see lines 145 and 225) but the overall form is acceptable
Experimental design: The study aims to correlate GADD45A expression obtained by IHC analysis to clinical pathological features. Compared to other studies it has the great advantage to present a large cases collection, patients’ follow up and survival information.

Material and Methods:
As better description of the IHC procedure (protocol and slides analysis) is now presented.
More importantly, the correlation between GADD45A and Ki67 was repeated considering the accepted cutoff of 20%, currently considered the reference value for this parameter. The revised analysis confirmed the previous findings.
Validity of the findings: In the first version of the manuscript the authors concluded that GADD45A was an independent prognostic factor for triple negative breast cancer, but this conclusion was not supported by a complete data analysis. As requested, statistical evaluation was repeated, including correlation with breast cancer subtypes and above all a multivariate analysis, that not confirmed the previous conclusion, therefore not providing the validation for GADD45A as independent biomarker for TNBC. Taking into account these results, the authors mitigated the emphasis on the clinical significance of GADD45A in TNBC. However, from this point of view the conclusion stated in the abstract regarding the possibility to use GADD45A expression level to stratify TNBC patients in risk categories, is probably still too speculative. In any case, all the requested revisions have been performed. Overall, a better description of the data and a more detailed and critical discussion of the results in respect of the state of the art was inserted, highlighting the different findings presented compared with similar studies, where opposite conclusion were shown. Considering a large patients' cohort analysed and the availability follow up data, the fact that authors were able to correlate GADD45A expression level with ER and PR status separately, this work could provide additional information that other investigators could take in to account.  The editor could decide if the findings are interesting enough for publication.
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: SUMMER DIATOM BLOOMS IN THE EASTERN NORTH PACIFIC GYRE INVESTIGATED WITH A LONG-ENDURANCE AUTONOMOUS SURFACE VEHICLE
Review round: 1
Reviewer: 1

Output:
Basic reporting: The paper is well-written. There are a lot of figures and supplemental materials but they all add to the story.
Experimental design: Given the intent (to deploy an autonomous vehicle to examine blooms in the SEP) the experimental design is well-justified. Autonomous platforms have inherent limitations, in that one cannot modify the data collection in real-time very easily, making the resulting analysis focus more heavily on data reduction and QA/QC. This is one of the main points of the paper--how do we use these platforms? I think the authors did a good job of providing both the pros and cons.
Validity of the findings: This manuscript describes a unique combination of autonomous platform and instruments. The authors have previous published on the use of the LRI Wave Glider platform for extended deployments, but that was with a more “traditional” suite of instruments. Here the inclusion of a LISST HOLO provides a unique new dimension. Key aspects of the paper cover both the technical strengths/limitations/issues with these platforms, and apply the data to examine questions about the importance of Hemiaulus in formation of blooms in the SEP.

The technical side of the manuscript provides a nice description of how these new platforms can be used, focusing in particular on data quality and the effect of fouling. The use of the LISST is quite intriguing, and the authors show how taking “bursts” of data can answer ecological questions while managing the considerable data volume produced. Again, the description of how the data were analyzed is useful in and of itself.

The science questions are a bit more fuzzy, with a lot of suggestions and directions for future research. For example, the authors speculate about the role of aggregation, and the potential for submesoscale interactions to promote blooms. This is all reasonable and interesting, but given the available data it must remain speculative. Despite that, some key conclusions emerge:

- there appears to be a background concentration of cells identified by the LISST
- early bloom formation is tied to small (regional) scale variability, and is very patchy—suggesting that autonomous observations are one way to get a better handle on the distribution and initiation of bloom events.
- aggregate formation is likely important

Taken together the combination of technical advances (included data reduction and analysis), and the more speculative assessment of the ecophysiology make this wirth publishing. There are a few specific items that should be cleaned up (see below). Beyond that, the one area that could be strengthened is the analysis of how cell density/aggregation is related to the physical data. It is suggested that ageostrophic flow may result in localized upwelling/downwelling, for example—is there any evidence for that in the other data (such as TS)? A more rigorous analysis would move the discussion into a more quantitative assessment of the role of small-scale variability.

Abstract: SEP is not defined

Lines 102-104: this seems a bit convoluted, in that blooms >10E4 cells/L extend well north at abundance up to 10E4 L-1. Is it necessary to give the abundance twice? And (minor point), greater than is not the same as up to. If this is listed twice because of conflicting reports as to the concentration, then rewrite.

118: add meters (2000-4000m)

154: the tether is variable in length, so it would perhaps be more appropriate to say for this particular configuration it was 7m.

183: not to be picky, but the configuration is based on both excitation and emission, so I imagine the LEDs (excitation) were also chosen for specific wavelengths, i.e. there was no white light source with filters.

382: While Honey Badger was the specific vehicle I suggest referencing WaveGliders more generically, since the emphasis is on what this class of vehicle can accomplish, not the specific vehicle.

Figure 4: the dashed lines are the SD, not the average. And remove the “v” at the end.

Figure5: define RA (running average?) and how you calculated it

supplemental information: shouldn’t there be specific figure captions for each of the supplemental figures?
Additional comments: I'd like to see this published, it is a unique dataset that demonstrates the potential of these platforms when some creativity in instrumentation and data analysis is used.


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

Input:
Title: SUMMER DIATOM BLOOMS IN THE EASTERN NORTH PACIFIC GYRE INVESTIGATED WITH A LONG-ENDURANCE AUTONOMOUS SURFACE VEHICLE
Review round: 1
Reviewer: 2

Output:
Basic reporting: See below
Experimental design: See below
Validity of the findings: See below
Additional comments: The manuscript by Anderson et al entitled “Summer diatom blooms in the eastern North Pacific gyre investigated with a long-endurance autonomous surface vehicle” (submission 28039) provides a descriptive account of spatial-temporal variations in phytoplankton properties in a region north of the Hawaiian Islands.  Data were collected from June to November 2015.  A particular interest in this study was the abundance of Hemiaulus and Rhizosolenia cells/aggregates.  The authors find these diatoms persistent in the study area and the occurrence of aggregates to span beyond the classic July-August summer export pulse (SEP) period, a finding which has implications on the role of these aggregates on carbon export and raises questions on whether aggregation is a consequence of physical processes or a normal life form.  I have to admit that I found this manuscript to be exceptionally well written and thorough.  The analyses are robust, limitations and challenges in the measurements are clearly articulated, and conclusions are soundly supported by the observations.

It is extremely rare for me to conduct a manuscript review and not have substantial comments to give, but in this case I have no concerns or criticisms and believe the manuscript is ready to proceed to publication.  I would like to congratulate the authors on a job 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: GOAL CONFLICT IN CHRONIC PAIN: DAY RECONSTRUCTION METHOD
Review round: 1
Reviewer: 1

Output:
Basic reporting: The manuscript is generally well prepared.  However, it does contain numerous errors such as number of participants not being equal to numbers approached minus those who refused!  In addition more work could be done to make it clearer why all the measures were used and to what purpose.
Experimental design: The design was a matched control group which seems appropriate.  However, it is not clear how well matched the groups are on variables other than sex, age and education.  Are they matched on paid employment, partner etc.

Otherwise the methods were well described.  Although it struck me as off that the conflicts measure did not seem to examine what the conflicts were between (e.g., work vs relationships, pain reduction vs exercise).  It was unclear to me how to classify a conflict that was between a goal in one goal category and a goal in a distinct category.  This needs to be made clearer.
Validity of the findings: The results are generally well presented and clear.

A report of which of the nine categories the goal conflicts were classified into would be useful.

It was also not clear why number of conflicts was focused on to be predicted by other variables in Table 3 or how this analysis was conducted.  Would it not be more interesting in the current context to examine predictors of the strength of the conflict?
Additional comments: This work explores some interesting issues although the value of the work is obscured by the way it is presented.  The authors could do more to highlight what are the key questions here and link these to specific analyses.  At present the manuscript is unclear and sometime confusing.  A revision should try to present a clearer, focuses manuscript and perhaps remove some of the less central analyses.


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

Input:
Title: GOAL CONFLICT IN CHRONIC PAIN: DAY RECONSTRUCTION METHOD
Review round: 1
Reviewer: 2

Output:
Basic reporting: The manuscript is unique because there is no previous, such detailed info about goal conflicts in chronic pain assessed with daily reconstruction method as well as there is lack of scientific publications about patients suffering from chronic pain and their daily conflicts.The introduction unit is very consistent and precisely describes the research.The manuscript does not need to be proofread by native speaker however inside abstract passive voice should be used more often than descriptive language as well as Authors should decide if they use numbers or words.There are about 57 cited publications including earlier ones and the latest. Most of the references are with impact factor.The results are clearly presented with figures and tables. The Manuscript is written according to the scientific manner with all needed structure inside.
Experimental design: The manuscript is original because  it fills the gap of knowledge about understanding of goal conflicts in patient with chronic pain. Research tasks are very well defined and clearly proceeded and all the results are meaningful.Experimental investigation is performed to a high technical and ethical standards.The methodology is fully compatible  with good scientific practice.The discussion is logical and rich in details. All the obtained results were described in details and discussed with available literature.
Validity of the findings: The manuscript is the novel publication about goal conflict in chronic pain. It is the very correct and reliable research manuscript describes the types of daily conflicts with the detailed analysis of its reasons and possible effects. All used data are harvest with the highest ethics and are analyzed in the very approved manner. Conclusions are well stated and linked directly to original research.
Additional comments: Dear Authors.Great job and I wish You all the further success and publications.


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: GOAL CONFLICT IN CHRONIC PAIN: DAY RECONSTRUCTION METHOD
Review round: 2
Reviewer: 1

Output:
Basic reporting: n/a
Experimental design: n/a
Validity of the findings: n/a
Additional comments: PeerJ #23277

The authors have generally done a good job of responding to reviewer’s comments and improving the manuscript.  However, a few issues remain.

1.	How were the 55 out of 181 control participants selected?
2.	Section 3.4.  Why report the data on numbers who did not report conflicts here rather than earlier before any data on conflicts are reported?  It would be useful to report n out of N alongside each percentage reported.  The description of the multi-level models was still unclear and was only understandable by consulting Table 3.  In addition, it seems problematic to look at each of these models as independent when the long list of dependent variables are probably strongly related (or at least not independent).  Perhaps this could be noted in the discussion.
3.	In the discussion I thought the authors could do more to note inter-dependencies.  Presumably if you come up with more conflicts about pain you probably mention less conflicts about other issues (given the number of conflicts people report is similar)?  Also how meaningful/useful is it to note that patients mention more conflicts at home and less at work if they are less likely to be in 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: GOAL CONFLICT IN CHRONIC PAIN: DAY RECONSTRUCTION METHOD
Review round: 2
Reviewer: 2

Output:
Basic reporting: The Manuscript is unique because there is no previous, such detailed info about goal conflicts in chronic pain assessed with daily reconstruction method as well as there is lack of scientific publications about patients suffering from chronic pain and their daily conflicts. The introduction unit is very consistent and precisely describes the research. The Manuscript was read by the native speaker that is why reading, understanding and text flow were improved. There are 57 cited publications including earlier ones and the latest. Most of the references come from impact factor journals. The Results are clearly presented and illustrated with tables and figures. The Manuscript is written according to the scientific manner with all needed structure inside.
Experimental design: The Manuscript is original because it fills the gap of knowledge about the understanding of goal conflicts in patients with chronic pain. Research tasks are very well defined and clearly proceeded and all the results are meaningfull. The experimental investigation is performed to a high technical and ethical standard. The methodology is fully compatible with good scientific practice. The discussion is logical and rich in details. All the obtained results were described in details and discussed wit available literature.
Validity of the findings: The Manuscript is the novel publication about goal conflict in chronic pain. It is very correct and reliable research manuscript and describes the types of daily conflicts with the detailed analysis of its reasons and possible effects. All used data are harvest with the highest ethics and are analyzed in the very approved manner. Conclusions are well stated and linked directly to original research.
Additional comments: After corrections it is even better and easier to read and follow. The whole text is very consistent. I wish all the team all the further successes.


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: PHYSIOLOGICAL AND BEHAVIORAL RESPONSE OF THE ASIAN SHORE CRAB, HEMIGRAPSUS SANGUINEUS, TO SALINITY: IMPLICATIONS FOR ESTUARINE DISTRIBUTION AND INVASION
Review round: 1
Reviewer: 1

Output:
Basic reporting: Overall, very persuasive presentation of a useful contribution to the field of invasion biology pertaining to this interesting species. There is some need for a little more basic editing for clarity and brevity. For example, the third sentence in the abstract (“We used an effective approach the…”)  seems to be missing a word or two somewhere in the middle and the first sentence in the introduction would seem to benefit from some reordering to place the subject at the beginning of the sentence. I recommend just re-reading or having an outside editor go over the ms one final time to do some tightening. To improve clarity and flow, I’d also recommend placing some of the inline citations more at the end of phrases or sentences, rather than in the middle of them (see Lines 56 and 58, for example).

The literature used seems sound and supportive, though I do worry that most of the work cited is from prior to 2010, so it may not reflect more recent work.

Abstract:
Content is informative and relatively clear, given some editing. Line 39 in the abstract may need to more clearly state that you’re referring to the rocky shore habitat, rather than ballast tanks.

Introduction:
Line 53 – should you say “… crab in the *rocky* intertidal…”?
Line 56 – As much work has been done on the invasion ecology of this species, I recommend including some more sources here – at least the review by Epifanio 2013.
Line 69-70 – Unclear inclusion. Are you intending to say that C. maenas is also estuarine, and thus a useful baseline for understanding? Need to clarify.
Line 74-78 – For brevity, I’d suggest just cite the 2003 review paper and the 2007 study, rather than provide the full list of cites and species?
Line 101-102 – I’m not sure what you’re saying here, but I don’t see that FW events are any more (or less) common along the eastern US than any other coastline – at least not without citation, and not sure the basis for the spring/fall pattern assertion. I think the important detail to make is that salinity can be highly variable, especially around estuaries, and that has a direct impact on the distribution of species.
L 103-107 – this sentence seems repetitive and tautological. You make a good case that we already understand that successful coastal invaders are often euryhaline (which already means they are good osmoregulators), so it’s unclear what the “need” is that you refer to. I’d suggest rewording this to, perhaps, say express that there is still some value in understanding this common characteristic further – or, more germane to your work, say that we still need to understand the role of behavioral choice, etc…
L 110 – I’m a little unclear on how behavioral preference is an indicator of risk. Isn’t it a measure of the ability of the animal to avoid risk? I may just not quite be understanding the use of the term “indicator” in this context.
L 112 – “…during ocean crossings,” needs a citation.
L 111-113 – This sentence seems to refer to the ability of H. sanguineus (larvae?) to withstand transport in ballast, etc., yet the rest of the paragraph discuss the current project – which doesn’t seem to be looking at ocean transport or larval survival, etc. It’s an interesting and important piece of information, but I’m not sure the sentence is relevant to the paragraph or the paper.
L 118 – “maladaptive” is not the right word to use here, given its typical use in a biological context. Perhaps consider something like “stressful” instead?
L 120-122 – Is this trait truly particularly important in decapods? I’m not sure what the basis is for that claim.
Overall, this sentence reads a little rough and vague. For example, what does it mean that they can alter environments? Perhaps consider revising.
L123-124 – “…as this is an a major…” remove “an.”
Experimental design: Experimental design seems reasonably well done and explained, though there is room for some editing for content and conciseness. However, I’m worried there may be some substantial errors in thinking. Particularly, the ballast water / Euryhalinity experiment has two major flaws as currently presented: 1) it uses adult crabs, whereas ballast water transports larvae, and 2) ballast water is not typically freshwater at all, at least not as experienced by this species. See Carlton 1985 and Bailey 2015 for a start to this topic. I’d suggest that all is far from lost, though – consider simply doing away with couching this in terms of ballast and focus on the (re)testing of the tolerance of *adult* H. sanguineus to different salinities.

Methods:
L 128-130 – some more details needed here: What size / age-class crabs? What were the acclimation conditions?
L 140 – I’d suggest not calling this “room” temperature. How variable was the “~”?
L 142 – How and how much were they fed?
L 144 – “Confirmation” of death? Maybe not necessary to state.

Overall, the statistical analyses seem appropriate, though I am not familiar enough with the specifics of some of them to fully comment. I do have a little concern about the application of multiple analyses to the same data, and the seeming post hoc nature of some of the tools used. But, I do like the relatively thorough and clear description of this portion of the methods.

The behavioral choice experiment is particularly nicely done.
Validity of the findings: Results:
I appreciate the brevity with which the results are presented, but some the specific results and their stats should be more expressly stated in the body of the section. However, the data does same appropriately robust and statistically sound.

L 238 – What does the word “suggested,” lease clearly state the results of the Kaplan Meier test here. In what way do these groups differ? Please state here.
L 244 – I’m not sure which analysis did not find significance in survival. Please state and, if appropriate, give the full statistical results in addition to the p-value.

Discussion:
Generally sound conclusions based on the present study, but much of the discussion is speculative and only loosely based on this and prior work. Again, I also strongly suggest some overall editing for flow and conciseness.
L313-315 – this is interesting speculation, but not clearly sound based on the current study. Please state more clearly.
L 332-334 – sentence unclear. Please reword.
L342 – “…may make it able to use of microhabitats…” missing word.
L343 – “…lesser…” should probably read “less.”
Additional comments: I think this paper makes a useful contribution to our overall understanding of this particular invader crab, and the role of salinity stress on shaping distribution. Given some reworking of some concepts (particularly regarding ballast water), a little clarification on some analyses, and some substantial style editing, this paper will be very worth of publication in PeerJ.

I look forward to seeing this work published.


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: PHYSIOLOGICAL AND BEHAVIORAL RESPONSE OF THE ASIAN SHORE CRAB, HEMIGRAPSUS SANGUINEUS, TO SALINITY: IMPLICATIONS FOR ESTUARINE DISTRIBUTION AND INVASION
Review round: 1
Reviewer: 2

Output:
Basic reporting: I feel there a bunch of areas where the grammar/descriptions can be tightened up or clarified. For instance:

In the abstract, "running trials" sounds like it could be "shrimp on a treadmill" or the process of conducting multiple trials.

line add "to" or something, e.g. "approach TO the salinity"

Line 36 I argue that "willingness" of crabs cannot be evaluated. Perhaps the authors could change to "capable"

Lines 57-60 can be shortened substantially.

Lines 74-81. I don't think ALL the species need to be listed. There are even some repeats. Perhaps just the authors.

Line 84 can be shortened to "H. sanguineus experiences salinity stress below 15 PSU..." "seawater" seems redundant.

Lines 108-111 can be shortened.

Lines 116-123. "This" is often vague, and it is used a bunch here. Also, I would prefer to see and end to the introduction dealing with the specifics of the study, not general statements about invasiveness. The salinity-invasiveness relationship should be mentioned earlier in the intro.

Lines 135-139 should be rearranged so grammar and logic are more inline.

Lines 146-149 are largely unnecessary.

Lines 153-164 - Right-sensored is not clear. Is it possible to briefly explain what this means? Are other references to sensored data then "left sensored"?

Lines 183-184 Beginning with "only two salinities" sounds like it is insufficient in some what. Do you mean that there is a convention of using 2 salinities? and you follow this convention?
Experimental design: I don't understand why the results in Figure 3 are all reported separately. It sounds like the experiment is a factorial design with male/female exposed to the different salinities and temps, but the data just show individual bar graphs of single factors. More importantly the analysis should look at interactions between factors if >1 factor was in an experiment. Some of these analytic  and design details are obscured by the often vague description in the methods.

Whys does table S1 only report female? This could be clarified in the text below S1.

Lines 153 - how many time of deaths were unknown?
Validity of the findings: Most of the discussion focuses on the osmolality across salinities, but most of the Methods/Results is about sex and acclimation differences. Can these be brought more inline?
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: PHYSIOLOGICAL AND BEHAVIORAL RESPONSE OF THE ASIAN SHORE CRAB, HEMIGRAPSUS SANGUINEUS, TO SALINITY: IMPLICATIONS FOR ESTUARINE DISTRIBUTION AND INVASION
Review round: 1
Reviewer: 3

Output:
Basic reporting: 1)	There a few places in the manuscript where statements are made without any supporting references.  Appropriate citations need to be added.  I have added “notes” to the PDF indicating these (ex. lines 112, 139, 334, and others noted in text).
2)	There are also several places in the manuscript where the meaning of certain sentences or phrases is unclear.  I have added “notes” to the PDF to point these out.
3)	The context for the study developed in the introduction and discussion focuses on the implications of salinity tolerance for promoting or preventing dispersal of organisms via ballast.  While the tolerance of adult crabs to salinity variation is highly informative for distribution potential within the invaded habitat, a more appropriate test of salinity tolerance would have to focus on larvae instead of adult crabs.  Because this study is focused on adult crabs, the authors should address the potential correlation between adult and juvenile/larval tolerance (using examples from literature), and should reduce the focus of the interpretation on this particular point.  Instead, the focus could shift to the role of salinity tolerance in the likelihood of distribution to or occurrence in different habitat types.
Experimental design: The number of samples used in experiments is quite good.  There are few areas where descriptions of methods should be elaborated or clarified:

4)	For the survival experiment, were crabs held individually or in groups?  If in groups, how was cannibalism (common in this species) prevented or accounted for as a cause of mortality for the survivorship study?
5)	The number of treatments that are reported is collapsed from the original 5 treatments into just 3 categories. The reason for changing the analysis and reporting seems to be (from wording at lines 171-173) that the analyses of each independent treatment did not show significant differences between survival from on treatment to another, so they were collapsed.  Is this interpretation correct, and if not, could this section be re-explained?
6)	Line 157-159 – what were the original set of parameters that were included in the models, before the final dependent variables were identified (ie. What was included in the models besides salinity?)
7)	The methods for the behavioral choice experiment do no include enough detail to be replicated.  The details of the trails themselves (how long a crab was monitored, how long it had to remain in its chamber or remain in the other chamber in order to be considered to have “chosen” those conditions) need to be explained.
8)	While the setup of the behavioral trials might allow for compelling evidence of the likelihood of movement out of the starting container, the approach does not appear to provide a signal allowing crabs to detect options outside of the chamber they are started in (as would be the case with a y-tube set-up).  This should be discussed.
9)	Lines 192-193 – this is the first time that a potential effect of temperature on salinity tolerance/avoidance is brought up.  A bit more context could be given here or in the introduction to explain why multiple temperature treatments are used.
10)	Lines 194-202 – How do temperatures of the behavioral choice containers compare to acclimation temperatures?  Were these set up so that crabs acclimated at 10 or 20C were in arenas that corresponded to these temperatures during the experiment?  The temperature set-up is no currently clear.  Line 201 also says salinity comparisons were varied between 5,15, and 30 – this sounds like salinities were changed?  The wording of this sentence should be changed for clearer interpretation.
Validity of the findings: These comments pertain particularly to the interpretation/discussion of the reported results:
11)	The while the first paragraphs of discussion do a good job of getting into the implications of the findings, there is little discussion or interpretation or several of the experimental results reported.  In particular, there is no discussion of the results of the behavioral assay:  males more likely to leave original container than females; effect of temperature on likelihood of leaving; preference for 35 PSU;  effect of acclimation salinity on likelihood of leaving container.
12)	The discussion also makes several statements about the relative competitive ability of H. sanguineus compared to other intertidal crabs, in terms of the possible advantage that H. sanguienus may gain due to their physiological salinity tolerance.  However, none of these statements are supported by comparisons to existing data about the salinity tolerances of other species with which H. sanguineus may interact.  I have listed a few relevant references below that the authors may wish to explore, in order to better inform (and potentially support) these statements.
13)	Lines 338-348 that expand on ideas of how H. sanguineus may exclude other crab species from the intertidal is interesting, but getting pretty unrelated to the results of the present study.  This information should be pared down in favor of adding additional discussion of the experimental results that are currently missing.
14)	The final couple of paragraphs of the discussion (lines 349-376) are quite broad, and provide some information that might be very helpful to cover in the introduction rather than the discussion.  For example, lines 363-366 succinctly introduce the idea that species vary in their salinity tolerances, and that these differences can structure species in space in estuaries.  This kind of information seems better suited to the introduction, perhaps in the first paragraph.
Additional comments: This study explores the effect of salinity acclimation on mortality and hemolymph osmolality of Hemigrapsus sanguineus, a non-native shore crab that has become highly abundant on rocky shores throughout the Western Atlantic from Chesapeake to southern Maine.  The authors also explore whether acclimation to different salinities influence the likelihood of crabs exiting their holding containers and whether the crabs end up in containers with different starting salinity.  The results of the study are interpreted in terms of how salinity tolerance may affect likelihood of H. sanguineus outcompeting other crab species, and surviving transit in ballast tanks.
The questions addressed in this study (whether H. sanguineus is behaviorally and physiologically adaptable in a way that may promote invasion success) is important, and the methods and level of replication are well-done.  The majority of my suggestions relate to the interpretation of the data collected, and to the context and clarity of its presentation.  I have provided general comments related to these concerns, and I have added additional notes directly to the review PDF with suggestions to improve clarity of the writing.  Overall, this is a meritorious study that will contribute to our ability to predict the current and potential distribution of this crab species.


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: PHYSIOLOGICAL AND BEHAVIORAL RESPONSE OF THE ASIAN SHORE CRAB, HEMIGRAPSUS SANGUINEUS, TO SALINITY: IMPLICATIONS FOR ESTUARINE DISTRIBUTION AND INVASION
Review round: 2
Reviewer: 1

Output:
Basic reporting: The overall clarity of the manuscript has been significantly improved, and I appreciate the authors attention to detail in addressing comments and suggested edits provided in the previous review.   There are a few remaining or newly introduced sentences that are slightly confusing, and I have added comments directly to the .doc version of the manuscript to point these out (apologies, but line comments were not possible as the line numbers re-start on each page, and no page numbers were included - something the authors may wish to attend to for future submissions).

The context for the paper has also been improved, with the new emphasis on the implications of wide salinity tolerance for geographic spread of established populations.
Experimental design: The clarity of methods reporting has also been much improved, including reporting of analyses used.  I have added a few additional comments on the .doc where ambiguities remain.
Validity of the findings: The reporting of results is also much clearer, and I appreciate the authors' more detailed discussion of the implications of the results.  A few minor comments are included in the manuscript.
Additional comments: The revised version of this manuscript coherently reports on new and sound data pertaining to the behavioral and physiological osmoregulatory capacity of an important invasive crab species.  The organization and clarity of this version are good, and the paper and should be considered for publication after correction of a few minor edits that will further clarify and streamline the text.


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: ROADS AFFECT THE SPATIAL STRUCTURE OF BUTTERFLY COMMUNITIES IN GRASSLAND PATCHES
Review round: 1
Reviewer: 1

Output:
Basic reporting: This manuscript would have benefited greatly from a native English speaker review before submission and I suggest review by a native English speaker during revision.  The text needs substantial revision to be clearly understood.

The manuscript describes some interesting work worthy of publication.  The authors have demonstrated that roadside habitat in Poland provide habitat for butterflies that is as good or better than reference sites.  Because of the road network, this habitat may serve not only as important habitat in and of itself, but also as a dispersal network.

However, the manuscript suffers from several issues.  The link between hypotheses/question, statistical tests, and conclusions is often weak.  The ms is heavy on statistical tests and weak on biological interpretation.  The reason for each statistical test should be clear, and the biological interpretation of each test should be clear.  The authors may want to consider removing some redundant or less informative tests to more clearly and carefully treat the remaining tests/hypotheses.  The introduction and discussion are quite short as a result.  Some elucidation of background and context would help the reader understand how generalizable the results are.

The figures seem to need axes labels.
Experimental design: This study is well within the aims and scope of the journal and is interesting work worthy of publication.

A figure of the transect design, or better text description, is needed.  The transect placement appears to be 5 parallel transects parallel to the road,  the first one 25m from the road, the next 25m further out, and so on.  But, it's not completely clear.

I couldn't understand the research questions on the first read through.  The knowledge gap is not stated explicitly.  The research questions are very broad.  The question of whether roads affect butterfly communities seems trivial, the question is what degree and how.  The reader would be interested in such things.

I have some concerns about the statistical approach.  For one analysis, the transects are treated as different, and for another analysis, they are lumped together.  That's not necessarily wrong, but the assumptions for statistical tests need to clear.  And the variety and number of multivariate tests has me concerned about spurious results.  The authors should be careful to identify mechanistic or biological reasons that can explain significant results, and if the results are difficult to explain, identify the results as a hypothesis to be tested further (McGarigal et al. 2000 Multivariate Statistics for Wildlife and Ecology Research).  There is little sense in the manuscript for the difference between descriptive/exploratory and inferential/confirmatory analysis and how much weight we can place on some of the results.  A significant p-value doesn't guarantee truth - there are well over 20 p-values in the ms, so a few are likely to be spurious.
Validity of the findings: Good practice is followed with treatment and reference sites.  Sample sizes are excellent.

Other than concerns stated above, I have concerns with overstating the importance of different butterfly species richness between road and reference sites.  38 species at reference sites vs 42 species at road sites, even if statistically significant, seems biologically very similar.  I wouldn't call such communities substantially different, unless there is something important about the 4 species missing from the reference sites.  However, it does seem important that roads have very similar, even almost identical, butterfly communities as reference sites, because one might hypothesize that roads would have negative effect on butterfly communities or populations.  The biology of the missing species might be quite interesting too, because it could provide hypotheses as to why roads might affect some species but not other species.

The argument that the results imply that roads are good dispersal corridors needs to be made carefully.  There is no data showing movement.  The argument seems to be that if roads are not affecting butterfly populations at roadsides, that they are then good for dispersal because of their distribution and connectivity on the landscape.

Some discussion is needed on what these population counts represent.  The reader doesn't have sense if these are butterflies passing through the sites or resident at the sites.  Is the surrounding landscape habitat for these butterflies?  Are the butterflies mostly confined to the sites?  So do population counts likely represent a closed, resident population?  Or are they a measure of butterflies that are passing through?  Are all the butterflies multi-voltine and reproducing for the entire period from April to September?  The reader needs some sense of the biology and landscape to be able to interpret what these counts mean.

In closing, I would like to reiterate that I find this work important and interesting and I look forward to seeing it published.
Additional comments: Please see additional comments in the attached pdf.


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: ROADS AFFECT THE SPATIAL STRUCTURE OF BUTTERFLY COMMUNITIES IN GRASSLAND PATCHES
Review round: 1
Reviewer: 2

Output:
Basic reporting: There are some grammatical inconsistencies throughout this article. I recommend that it is edited carefully to improve English grammar.
Experimental design: I think the experimental design as it pertains to the butterfly data is very novel. The research fills knowledge gaps of butterfly communities on road verges. I do think that the plant community data as some limitations which are not currently noted in the manuscript; namely only one sample was made during the growing season. In North America, one vegetation sample of cover would not adequately measure plant communities because some would have senesced by then.
Validity of the findings: The data seems sound and the benefits to the literature are clearly stated.
Additional comments: I'd like to see you address roadside management in the methods section as it pertains to the road verges in the study, and a brief overview in the discussion, since management has a huge influence on butterflies.

Thanks for your work on this study!


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

Input:
Title: ROADS AFFECT THE SPATIAL STRUCTURE OF BUTTERFLY COMMUNITIES IN GRASSLAND PATCHES
Review round: 2
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: Article is much improved and very interesting to read.


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

Input:
Title: TRICHOCERA MACULIPENNIS (DIPTERA)—AN INVASIVE SPECIES IN MARITIME ANTARCTICA
Review round: 1
Reviewer: 1

Output:
Basic reporting: Review of “Trichocera maculipennis (Diptera) – a potential invasive species in maritime Antarctica” for PeerJ.
By Potocka and Krzeminska
This mss, which is long at nearly 450 lines with four plates of figures, documents a new location for an exotic fly on King George Island.   The supposedly new information shows that the species can spread to scientific stations across glacial barriers.  The manuscript also includes several pages on the systematics, life history and distribution of the species which has already been published by the second author with other authors (2013).   Moreover, earlier documents, including a publically available Working Paper (CEP 10a, 2016) have already documented the new locations and its spread is not surprising as it has already been recorded outside station buildings in the natural environment.  The inclusion of figures 3 and 4 are also unnecessary as all of them have been published before by  Krzeminska.  I do not consider therefore that the publication of this paper is justified in its current form as it only records a new locality for the fly and does not advance scientific knowledge of the species.
What I would have liked to have seen in this mss was a discussion of the attempts at eradication including details of methods used, why they failed, how the method could be improved by paying more attention to the vulnerability or not  of different stages in the species’ life history.   For instance,develop an eradication plan  using not one method, but several different methods could be used, each targeting a different life stage.   Information from attempts to eradicate a different species from the Australian Casey Station is relevant but is not mentioned.  To make space for the eradication discussion I have marked on the text which sections should be deleted.
My detailed comments are below.
•	The English is poor and clearly has not been checked by a native English speaker.  Practically every manuscript I am sent to edit is authored by non-native English speakers who have not bothered to have their writing edited.  This wastes a deal of my and other referees and editors’ times and is quite annoying.  This mss is one such and although I have taken time to improve the text, there is more to be done.  I urge the authors to elicit the help of someone or several people who know and have experience in writing scientific papers in well crafted English before submitting again.
•	 There are numerous omissions in particular any references to the exotic mycocelid fly in the sewage treatment plant (tanks and pipes) at Casey Station and authors seem ignorant of it (line 318).  This fly was eventually eliminated I beleive, although initial attempts by engineers failed while an entomologist was consulted  but their recommendations not followed.  The problem at Casey was that the pupa was resistant to heat, cold, moisture, chemicals etc etc and hid in nooks and crannies in the sewage system.  The authors do not mention this possibility  with T. maculipennis pupae nor do they suggest that extermination is necessary and discuss methods (see above).  Admittedly they recommend an action plan be prepared jointly by four countries but they do not suggest any method of removal nor any plan as to how these four nations will be able to produce one.  I can see this being a ten year project meanwhile the fly expands its range.  Control and extermination should start now.
•	The authors seem convinced that the fly has colonised the additional stations by long term natural dispersal but I suggest this is unlikely.  Despite the fly being apparently immobile in strong winds, there is no reason to suppose it could not have been dispersed on wind currents accidentally.  The authors seem not to have considered this possibility but must do so and include in the paper the routes across and along the coast of the wind at different times of year.
•	What is more likely by looking at the current records being only at scientific stations, is that it was human agency that dispersed the propagules.  The authors do not document whether there was any human traffic between Stations but I suspect that there is and was. What about tourists for instance and “adventurers”.  It could well be difficult to get this data but it must be added.  Again this route is not considered by the authors .
•	In the title, I do not know why the authors consider the species a potential invasive.  Surely the new dispersal records show that it is invasive.  Has it been documented as invasive in other parts of the world?
•	Why were specimens not bar coded?  Specimens from Kerguelen should also be barcoded as well as from each station on King George Island.  I would be surprised if there are not barcodes from European specimens in GenBank for comparison.  The origin of each population could then possibly be established.
•	It is well known that fresh fruit and vegetables imported into the Australian Antarctic and subantarctic Stations are the source of introductions of exotic invertebrates.  Indeed, some stations have banned the importation of these commodities for this reason.  The mytocelid fly at Casey is a pest of mushrooms for instance. So the best method of preventing reinfestation, if it is eliminated, is not to import fresh root vegetables in particular.  That will be cheaper in the long run than a succession of eradication efforts.
•	The life history of the fly is described in detail much of which is not relevant to the situation on King George Island. Instead, information is needed on the conditions in the sewage plant, i.e. temperature and the tolerance of the species to polluted waters and especially if the pupa is hardy and cryptic (see above). Some information only is given of its behaviour in non Arctic locations.
•	n dashes and not hyphens should be used for all spans i.e. page numbers. All references are given in the wrong format.
•	The figures 1 and 2  are useful and of a sufficiently good standard to be included including a complete rewrite is needed with attention to all my comments above.
Experimental design: See above.  n.a.
Validity of the findings: No findings - see above
Additional comments: See above.   Total rewrite necessary and collection of more data.