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: MITOCHONDRIAL GENOMES OF THREE TETRIGOIDEA SPECIES AND PHYLOGENY OF TETRIGOIDEA
Review round: 1
Reviewer: 3

Output:
Basic reporting: The authors present the description of three mitogenomes of Orthoptera of the family Tetrigidae, basically in a data release manuscript. The manuscript is well written and easy to understand. The structure is adequate, as the results are rather descriptive and leave comparably little room for discussion.
Experimental design: The methods are suitably applied, largely well described, and the results are described in high detail.
Validity of the findings: Overall, the findings appear valid. My main point of criticism is the following:

Molecular phylogenetic studies of Caelifera are notorious for the difficulties presented by mitochondrial haplotype sharing (see references pp. 6/7 in Hawlitschek et al. 2016, http://onlinelibrary.wiley.com/doi/10.1111/1755-0998.12638/full). While most previous studies focus on Acrididae, not on Tetrigidae (or related families in Tetrigoidea), the authors should take into account that hybridization (which has been postulated also across genera) or other phenomena causing misleading results may also influence their Tetrigoidea results. The authors should introduce these problems and carefully discuss their implication for their results.
Additional comments: Other than that, I only have minor comments, a list of which is given below.

Kind regards,

Oliver Hawlitschek

l. 16: Replace "vaule" with "value".

l. 30 / 31: Add citations for the first two sentences of the introduction.

l. 39: "minor damage to crops": better write "minor importance as agricultural pests".

l. 47: "original taxon": does this mean "sister group to all other Tetrigoidea"?

l. 53:  Replace "widely used molecular components" with "a widely used molecular component".

l. 69: "PCR amplification and sequencing by primer walking": Text should be bold.

l. 89 / 90: "18 members of Caelifera and F. qinlingensis, […]": But the species of Tetrigoidea studied here are members of Caelifera, too?

l. 128: "positive relativity": Do you mean "positive correlation"?

l. 132: I do not understand the importance of true bug mitogenomes here. If this is a standard method I do not know it. Please explain in the methods section.

l. 172-174: "Some highest […] encoded tRNAs.": These sentences are difficult to understand, please check the wording.

l. 224: "original taxon": See my comment to l. 47.

l. 230: Replace "Melanopline" with "Melanoplinae".

l. 237/238: "topologies of Tetrigoidea based on 16S rRNA": It is not clear where these results come from. Did the authors conduct specific analyses based on 16S only? This is not described in the methods section. Or are they from Eades et al. (2014)? Please clarify.

l. 270: Replace "Zhemin zheng" with "Zhemin Zheng".

l. 393: Replace "Potua Sabulosa" with "Potua sabulosa".

l. 397: Please give the website for the download of Tracer.

l. 406: Replace "Molecular evolution" with "molecular evolution".

l. 422: Italicize "Tetrix tenuicornis".

l. 440: Replace "Comparative & phylogentic" with "comparative & phylogenetic".


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: MITOCHONDRIAL GENOMES OF THREE TETRIGOIDEA SPECIES AND PHYLOGENY OF TETRIGOIDEA
Review round: 1
Reviewer: 4

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: There is one aspect in the paper which I find speculative at the moment. I explained it in a comment in the attached pdf (line 150).
Additional comments: I added some comments and suggestions 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: MITOCHONDRIAL GENOMES OF THREE TETRIGOIDEA SPECIES AND PHYLOGENY OF TETRIGOIDEA
Review round: 2
Reviewer: 1

Output:
Basic reporting: Following on my previous comments:

Section 'Nucleotide composition and skew':
The phrase strating with 'The approximate positive correlation were found (...).' need revision. A-skew and C-skew should be replace by A+T-skew and G+C-skew for the sake of consistency.

All figures still need to be improved with regards to resolution for publication. For example, I found it difficult to read a printed version of the phylogenetic tree.
Experimental design: No comment.
Validity of the findings: No comment.
Additional comments: No additional comments


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

Input:
Title: MITOCHONDRIAL GENOMES OF THREE TETRIGOIDEA SPECIES AND PHYLOGENY OF TETRIGOIDEA
Review round: 2
Reviewer: 2

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


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

Input:
Title: MITOCHONDRIAL GENOMES OF THREE TETRIGOIDEA SPECIES AND PHYLOGENY OF TETRIGOIDEA
Review round: 2
Reviewer: 3

Output:
Basic reporting: No further comments.
Experimental design: No further comments.
Validity of the findings: No further comments.
Additional comments: The authors have thoroughly revised their paper, improving it considerably. I only have a few minor details, mostly stylistic, that should be taken into account.

Kind regards

Oliver Hawlitschek

1. l. 33: "and one tribe": Xerophyllini is a tribe within Cladonotinae. Other subfamilies of Tetrigidae have more tribes. Best remove this.

2. l. 53: Change "Cong and Grishin., 2016" to "Cong and Grishin, 2016".

3. l. 59: Change "represent for Scelimeninae, make it possible" to "to represent Scelimeninae, making it possible".

4. l. 60/61: Change "Combined with other 18 Caelifera mitogenomes in the database, the comparative analysis was made" to "A comparative analysis with other 18 Caelifera mitogenomes was conducted". But see my next comment.

5. l. 54-66: Most of this entire paragraph is a summary of the methods section and therefore redundant. I suggest keeping only the first, second, and last sentence and removing the rest of the paragraph.

6. l. 156: Change "the identify of Tetrigoidea" to "the identification of Tetrigoidea".

7. l. 176: Change "NNU were relatively" to "NNU was relatively" and "NNC were lower" to "NNC was lower".

8. l. 178-182: "Some highest … tRNAs": This sentence is still not clear. Please rephrase again.

9. l. 245: Hawlitschek et al. (2016) is not the ideal reference here, as it does not really discuss hybridization. Check p. 2 of this paper for more suitable references (e.g., Gottsberger 2007, Hochkirch & Lemke 2011, Rohde et al. 2015).


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 FURVELA TENT-TRAP MK 1.1 FOR THE COLLECTION OF OUTDOOR BITING MOSQUITOES
Review round: 2
Reviewer: 1

Output:
Basic reporting: This manuscript “The Furvela tent-trap Mk 1.1 for the collection of outdoor biting mosquitoes” by Charlwood et al. describes the refinement of the Furvela trap for the collection of anthropophilic mosquitoes. To analyze the new configuration of the Furvela trap for endophagic biters, they compared twice-a-week usage of the tent to daily window-trap collections in Northern Tanzania, and for exophagic biters to outdoor light traps in the PAMVERC trial area in Mozambique. Overall, I believe the trap design to be a good one, especially due to the flexibility of the general design, cost, and due to the ubiquitous nature of CDC light traps.  I do think the manuscript suffers from not including more from the large PAMVERC trial data as this has hundreds or thousands of trap nights, and from ambiguity in the methodology.

I fully agree with the premise that human landing catch is becoming unacceptable for use, and its use needs to be phased out. With this, I do feel that a more direct comparison between the Furvela design and HLC should be done. I do not believe this is necessary for publication, but I do think it is necessary to a widespread adoption of the technique.

I do recommend the paper for publication, though think a major revision of the text should be undertaken to improve clarity for the reader. I also feel the format and quality of the figures needs to be improved prior to publication.

Basic Reporting:
The Furvela trap, while previously described, needs to have a basic description in the text to orient the reader to the design. This is especially pertinent as I can’t locate the (what I presume) is the original description of the Furvela trap, as this reference (Charlwood JD: A new efficient and cheap trap for the collection of outdoor-biting mosquitoes. Acta Trop 2005, 95:S1–S506) seems to be to the entire supplementary info covering conference proceedings from the 4th MIM Pan-African Malaria Conference. The supplementary video and SOP do help this, but still I feel the paper would benefit from something brief in the text before the “improvements” part of “The Furvela tent-trap Mk 1.1” section.

Figures as included have the below issues:
•	They may not be of sufficient resolution.
•	Figure 1 Legend: I would highly recommend listing in the legend what each picture is showing. Defaulting to the text makes description unnecessarily taxing.
•	Figure 2 Legend: “Tanzania” is misspelt.
•	Figure 2: I don't see an arrow.
•	Figure 2: Why is only the blue line labeled?
•	Figure 2: The gaps in sampling would be better presented with a broken axis as you then lose information from the time periods you have sampling (i.e. reduce the “white space” in the figure).
•	Figures 3 and 4: I'd highly recommend the use of bar graphs over pie charts. It would convey the data in a smaller and more informative package. Additionally, this allows for easier comparison between groups. Stacked barplots may make comparisons between before/after easier. See below figure (in attached pdf) for example (I will also attach the R code to generate this figure). This presentation also allows you to more quickly look between species without having to go back to the figure legend to see what is what.

(see attached PDF for figure)

•	Figures 3-4: Make sure that colors are consistent between graphs, i.e. Furvela is always blue.
•	Figure 4 Legend: A numbered reference is cited, but the references aren’t numbered.
•	Table 2 uses “DRR” which is not defined. The text describes “IRR.”

Other comments:
•	Lines 206-207, Where is this time data? If in the supplemental I only see times "1, 2, 3". Was the night divided for collection period? This needs to be said explicitly how the night was split if so.
•	Overall, the supplemental data could be cleaned up significantly. i.e. in the indoor_outdoor_furvela document, there are extraneous datasheets that aren’t included in the paper, sheets are labelled poorly (“Sheet 1”, “Sheet 2”, “Collections”, etc..), and some of them have no/poor labeling (no dates, shorthand for species). Fix sheet labels, remove datasets not utilized, and remove figures not used.
•	The supplementary file names need to be explicit and the same as described in text, i.e. include “Supplementary File 1 – xxxxxxxx.xlsx”.
Experimental design: The design is significantly muddled by the cobbling together of various studies using the Furvela trap, of which it is poorly described which tents are used (models, etc.), which “improvements” are included, and sampling locations/times/methods compared, etc. I think the authors would benefit from a descriptive table of what sampling was performed.

	The methodology needs to be expanded, i.e. number of trap nights, be explicit about what data is re-used from studies published elsewhere, design of the study (i.e. are collectors being rotated through tents, etc.?).
Validity of the findings: A central premise regarding the furvela trap’s uniqueness is that an “entry” behavior is not required for collection. However, as mosquitoes are caught while attempting to enter the hole in the tent (while following the odor plume)---I feel this distinction is subtle from other “entrance” traps. The design is like that of the “infoscitex tent” design we described in Krajacich et al. as mosquitoes collected to odor plumes pushed out of mesh windows are instead collected by suction ports above the mesh (albeit in a simpler design with the Furvela trap).

Mosquitoes have been known to be very adept flyers, entering even small holes in a bed net. While the fan I can imagine makes entry difficult, as there is a large gap in the zipper- how often have collectors found mosquitoes in the tent? How often have you caught full or partially bloodfed mosquitoes in the furvela trap (indicating they were possibly caught while exiting after feeding), and does this proportion differ from that caught in the CDC Light traps?
Additional comments: Line 147: Need a comma after “light-traps”
Typo in line 151, “In Kagera Province, northern Tanzania.” should not be a separate sentence.
Line 214: Remove comma after "In studies"
Lines 224-226: Is it true that only An. funestus and Culex spp. have changed indoor/outdoor ratios? It seems only Mansonia africanus is the same before/after spraying. The two mentioned are the only ones who went from dominant indoor to dominant outdoor, but most changed in proportion. The discussion should address whether any of the vectors (especially those such as Coquelletida fuscopenata that are less-commonly described) have been reported to be more commonly exophagic in other studies (from indoor/outdoor light traps or ideally HLC).

Line 249, need comma between "outdoors" and "suitable"

Line 254: "Window-traps may be a substitute for light-traps." What is this based on? They may be for indoor collections, but obviously not for outdoor collection.

Lines 279-280: Has this light repellence been seen with CDC light-traps with/without the bulb (but having a CO2 source) in this area? This finding is interesting as I am unaware of a documented repellence effect due to light for anthropophilic vectors, though I am aware of many cases of lights attracting vectors (i.e. sandflies) and other non-target dipteran (making sorting more difficult).


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 FURVELA TENT-TRAP MK 1.1 FOR THE COLLECTION OF OUTDOOR BITING MOSQUITOES
Review round: 2
Reviewer: 2

Output:
Basic reporting: The manuscript "The Furvela tent-trap Mk 1.1 for the collection of outdoor biting mosquitoes (#16725)" describes a modified version of the Furvela tent-trap and compare it to other trapping methods.

The English language requires significantly improvement, e.g. have to be checked by a native speaker. I have indicated several sentences which stay unclear for me due to language and grammar issues.

Although the general structure of the manuscript conforms to PeerJ standards, the structure of the introduction is not very easy to follow. Please start with the general background (malaria, number of cases, distribution, ...), then explain the Motivation of your study (indoor collections to not necessarily reflect the outdoor situation, ...) followed by the State-of-the-art (pros and cons of methods used in the past (HLC) and current methods (light traps, tent traps)). Then highlight the lack of knowledge (there are already tent traps available, explain why we need your improved system (entry behavior, ...)) and finally give a description of the explicit study aims (in short: What exactly are you doing and why). In addition, several information are not well referenced.

The figures and tables are generally okay. However, I recommend using bar plots instead of pie charts, which are generally expected to be more comparable between different groups. Some information are missing in the labels of the figure and tables, e.g. explain all abbreviations in table 2 in the label (DDR, TT, CI, adjusted, ...).

Finally, although the raw data are supplied, the data are not very useful. Every table has a different, mostly unstructured format and use different abbreviations, which are not explained.
Experimental design: The research is in the scope of PeerJ, but the explicit knowledge gap and research question is not well described.

As highlighted for the introduction, my critique of missing structure also applies to the methods section. You are comparing the Furvela tent-trap in different settings, but due to the unstructured presentation, it does not become clear what you did in which site. Please give different subsections, e.g. "In the first experiment, we did ..." &  "The second experiment was conducted....", etc. In addition, from the description in the methods, it does not completely becomes clear, that the monitoring was conducted after mosquito control interventions. Furthermore, important information on the mosquito sampling (e.g. window traps) and statistical methods (e.g. why negative binomial) are missing.
Validity of the findings: The findings are valid and supported by sufficient data, but are not presented very well. As in the introduction, there is a lack of references for some of the statements and therefore sometimes look like speculation. The research question was not clearly indicated and therefore there is no link between the introduction and the conclusions.
Additional comments: INTRODUCTION
#before line 39: please provide general information on the impact of malaria on public health in Africa (number of cases, distribution, etc.)
#lines 40-42: "This success ..." --> unclear sentence, please rephrase.
#lines 42-44: "Outdoor transmission [...]" --> unclear sentence, please explicitly explain why both, indoor AND outdoor monitoring, is needed, i.e., because indoor monitoring does not necessarily reflect the outdoor situation
#lines 44-50: "Indeed, ..." --> I do not understand why you include this citation. Delete?
#line 52: "untested," --> unnecessary comma
#line 55: "require" --> do you mean "lead to"?
#lines 56-58: "Although ..." --> Please be more specific: malaria vectors = Anopheles spp. or specific Anopheles species (Anopheles gambiae complex). In addition, the vectors for malaria can also transmit other pathogens.
#lines 59-61: "Outdoor ..." --> Please give references for the statement that human landing collections is the best method.
#lines 61-62: "HLC ..." --> Please give a reference.
#lines 62-63: "Such ..." --> Please give a reference.
#lines 63-67: "The objective ..." --> Unclear. As far as I understood, here you want to explain why alternative methods for HLC are needed.
#line 70: What do you mean by "risk-free"?
#line 72: What do you mean by "at least as far as malaria vectors go"?
#line 74: At the first report of a species in the manuscript, please give the full name of the species, e.g. "Anopheles gambiae s.l." instead of "An. gambiae s.l."
#line 75: replace dot by comma: "CDC light-traps." --> "CDC light-traps,"
#lines 93-94: "The ‘Furvela’ tent-trap, ..." --> Why does it not require an entry behavior? Is there a reference of the first description of this trap?
#lines 94-95: "The trap remains ..." --> reference?
#lines 110-112: "We supply ..." --> Was the aim of the study to show that the Furvela trap catach mosquitoes? I thought this was clear before. What exactly are the aims of your study?

METHODS
#before line 116: please give some more information as introduction of the method section, e.g. "in order to improve the Furvela tent-trap Mk 1.1 ..." or information why the improvements were necessary
#line 116: please give a reference for the first description of the Furvela tent-trap Mk 1.1
#line 147: include comma after "In addition to CDC light-traps"
#lines 147- : I completely missing details on the exit window-traps (e.g. construction, references, etc.).
#line 154: "The best ..." --> What trap do you mean? Window-traps or Furvela traps?
#lines 155-156: "As mentioned ..." --> As mentioned before? You have to explain, which risk a linked to HLC and give a reference for the statement.
#line 158: What is "PAMVERC trial"? Explain.
#lines 165-167: Why did you use negative binomial regression, e.g. Why did you not use ANOVA or Poisson regression or .... Please explain how you observed overdisperion in the data, which is one important information to select negative binomial regression as appropriate method. Which explanatory variables were used? Was this statistical method also used to analyze the collected data described in the lines 178-183.

RESULTS
#line 203: comma after "Between the 23/10/2014 and 14/3/2015"
#lines 205-206: "r = 0.93, P = 0.0003" --> What kind of statistical method was applied here? Spearman? There is no explanation here or in the method section.
#lines 206-208: "A larger proportion ..." --> As far as I understood from the method section, you only aimed to compare the different kind of traps? Why did you also analyze the temporal pattern? Explain in the description of the study aims in the introduction and explain in the methods how you analyzed the temporal pattern.
# lines 208-212: "Immediately following the application ..." --> Please explain this mosquito control measurements in the method section and not in the results section.
# lines 214-216: "In studies, from Tanzania, ..." --> Please move to the discussion section.
# lines 216-217: "The other species ..." --> Which other species? Describe.
# lines 217-219: "Comparable results were ..." --> What do you mean by "comparable results"? Describe.
# lines 233-234: "The incident rate ratio ..." --> I do not understand what you are doing here. Give more information on the IRR and the variables TT and L.
# lines 234-236: "Variables including collector, ..." --> The modelling approach must be explained in the method section.
# lines 234-236: "Variables including collector, ..." --> What do you mean by the variable "collector". Might be better included as a random effect?
# line 239: "Anopheles" in italics

DISCUSSION
#general: Please also discuss potential problems with the tent traps. For example, please discuss the problems of the sampling standardization, which might be highly dependent on the collector in the tent.
# lines 247-248: "Because of the risks ..." --> Give a reference.
# lines 258-249: "In order to determine ..." --> You do not determine the transmission, but the density of potential vectors. Please rephrase.
# lines 249-252: "The ratios ..." --> In general, the comparison does not give any information about the suitability of the Furvela tent-trap to trap outdoor biting mosquitoes, e.g. because you do not have information about the size of the real outdoor biting mosquito population, but indicate a similar performance like the light-trap. Please rephrase.
line 254: "Window-traps may be a substitute for light-traps." --> What do you mean by "substitute"? I do not understand.
#line 258: "sp." --> not in italics. In addition, please use "spp." as there are several species present in Cameroon.
#lines 267-268: What do you mean by "chaotic"? Spatial-temporal variability? Please explain and rephrase.
#line 269: "indoor light-trap" instead of "light-trap"
#line 269: "For example" instead of "Thus"
# line 277: "With the current ..." --> You mean the elimination of mosquitoes in order to eliminate malaria. Rephrase.
# lines 285-288: "Ease of transportation ..." --> Why are vector maps needed? Explain.
# lines 288-289: "Such information ..." --> How do the tent traps help to estimate the flight range of mosquitoes? Completely unclear for me. Explain.
# lines 289-290: "They may ..." --> How to tent traps help in the regarded to determine the intervention area? Explain.
# lines 307-308: "Presently this ..." --> Why is there a restriction to indoor insects? Explain.
# line 308: "The use ..." --> I do not understand the sentence. Rephrase.


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 FURVELA TENT-TRAP MK 1.1 FOR THE COLLECTION OF OUTDOOR BITING MOSQUITOES
Review round: 3
Reviewer: 1

Output:
Basic reporting: I feel the manuscript has been greatly improved since the initial version, both in terms of clarity and the overall formatting. I feel that the changes that have been incorporated are appropriate and sufficient.

Below I list some minor corrections/suggestions.
Lines 36-39: This sentence is a bit awkward/run-on and would be better split into two. Also merge into below paragraph.
Line 239: Report week numbers or change figure to show dates
Line 242: Report density drop
Line 278: Remove “really”
Line 319-321: I’d mention explicitly that light being a repellent has not been shown previously.

Overall comment: The discussion is somewhat disjointed and doesn’t flow easily from paragraph to paragraph. I’d suggest some improvement of transition sentences between paragraphs, and the paragraph order. I think it’s publishable in this format, but could be improved.
Experimental design: no additional comment
Validity of the findings: no additional comment
Additional comments: No additional comments


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

Input:
Title: THE FURVELA TENT-TRAP MK 1.1 FOR THE COLLECTION OF OUTDOOR BITING MOSQUITOES
Review round: 3
Reviewer: 2

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The authors significantly improved the manuscript and can be accepted. However, to my opinion, the supplementary tables still need significant improvement. Especially please do not use unexplained abbrevations (e.g. "coll" = collection method, "afr" = "Anopheles funestus", AG part = ???, ODDS = ???).  Please tidy these tables up, there are a lot of blank cells or unused columns.


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: NORTH ATLANTIC OSCILLATION DRIVES THE ANNUAL OCCURRENCE OF AN ISOLATED, PERIPHERAL POPULATION OF THE BROWN SEAWEED FUCUS GUIRYI IN THE WESTERN MEDITERRANEAN SEA
Review round: 1
Reviewer: 1

Output:
Basic reporting: I outline specific issues to be fixed below:

-This manuscript uses clear language throughout, although it would benefit from an additional readthrough by a native English speaker.

-The last paragraph of the introduction should be removed -- it is more suitable for inclusion in the abstract, or perhaps the discussion.

-In lines 160-161, the authors state that the 'AO is characterised by a merdional dipole in sea level atmospheric pressure between polar regions and mid-latitude'.  This is the NAO, not the AO.  The latter half of the sentence  '...and could be interpreted as the surface signature of modulations in the strength of the polar vortex aloft' is verbatim from the abstract of Thompson & Wallace 1998 and should be rephrased.

-Throughout this manuscript, the authors write out and explain equations that are standard in the field. (A particular example of this is the approx. 1 page devoted to explaining chlorophyll fluorescence on lines 271-289.) Replacing this text with one-two sentences and a citation for chlorophyll fluorescence would suffice. Similarly, equation 1 and the text following (lines 127-131), equation 2 and the lines following (137-142), etc. could be removed without losing any clarity in the manuscript.

-In the figures (3, 5, 6) please use different shaped symbols in addition to different colors to distinguish among your results. This is necessary for any color-blind readers, as well as printing in black&white.

-In figure 4, please clarify what the grey and orange boxes represent. And, please make the shading/hatching different between the two colors (see color blind readers, above).
Experimental design: The method of sampling for Fucus guiryi presence/absence needs to be described in the Methods: how was it checked? Quadrats and transects? Visual searching over a defined period of time?
Validity of the findings: The most significant issues with this manuscript are the assumptions made about the basic biology of Fucus guiryi, which then impacts the interpretation of the authors' findings in the abstract, conclusions, etc.  The authors assume that the presence of F. guiryi in the summertime is solely due to the transport of thalli and embryos from neighboring populations to the west.   However, it is always present at their key study site in the winter. (It is not stated how the winter populations appear.) No mention is made of the reproduction of F. guiryi and the possibility of microscopic stages/remnant holdfasts of macroscopic stages remaining even when juvenile or adult thalli are not present (and thus it appears to the naked eye that F. guiryi is gone).  The authors could have tested specific issue for this by putting out settlement plates at their sites during different years -- any F. guiryi that appeared during a summer season would have to be due to the settlement of new individuals. Also, the fact that F. guiryi was present _every_ winter of the survey indicates the low likelihood that this population is solely dependent on recruitment from a western source population.  In a broader context, it is possible that the appearance of summer F. guiryi is correlated with the NAO...not because the NAO brings F. guiryi to this site, but instead because it creates favorable conditions for existing F. guiryi to grow into macroscopic sizes.  The authors should consult Schiel and Foster 2006  doi:10.1146/annurev.ecolsys.37.091305.110251 for a review of the population biology of fucoids.

The authors state that Tarifa is the 'seeding' population. As this was not conclusively demonstrated/experimentally tested, they should state instead that it is a 'likely seeding' population. (see comments in #1 above)

The authors do not examine or discuss the impact of herbivores on blade shape, which can significantly impact their discussion/results of developmental instability. Some discussion of the impacts of herbivory on blade shape is needed.
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: NORTH ATLANTIC OSCILLATION DRIVES THE ANNUAL OCCURRENCE OF AN ISOLATED, PERIPHERAL POPULATION OF THE BROWN SEAWEED FUCUS GUIRYI IN THE WESTERN MEDITERRANEAN SEA
Review round: 1
Reviewer: 2

Output:
Basic reporting: MAIN: This paper reports a long (26 years, 1990-2015) time series of records of occurrence of a marine alga at an isolated warm range limit. This is a fantastic opportunity as such a data set is difficult to obtain and has allowed the authors to make a more detailed analysis of causes of variability in presence / absence than what is usually possible.  The analyses conducted allowed a correlation of the presence of the species with the overall mean value of the NAO (an index reflected in mean weather conditions) from the spring months.  Spring is the main reproductive / recruitment season for the species, This is an important topic that deserves publication. I recommend that it should be published quickly.

Main suggestion 1. The paper could be even further improved if the authors could try to use a better proxy for the local conditions of this very localized study area rather than just the NAO.

Main suggestion 2. The introduction could be more clear in the formulation of hypotheses/questions behind the other measurements conducted.  The measurements conducted in July 2011 (radiation, temperature, photosynthetic parameters) are a single point in time relative to the time series and it is hard to related the two things and to understand the aim of these point measurements relative to the goal of the paper. I recommend a better explanation of what question are you trying to answer with these measurements.  Also, a comparison conducted in the field, under distinct environments can be due to population differences or environmental differences. So, again, please explain better what is the question.

Main suggestion 3. I recommend a better literature review of other such cases of variability of edge populations of Iberian algae.

3a. First, please compare your results with the very relevant paper on variability in Fucus guiryi at its southern limit by Carla Lourenço et al (2016). It reports the dependence of this species on specific local climatic effects namely the upwelling conditions in isolated spots.  It also reports the southern limit of Fucus guiryi.
Lourenço CR, Zardi GI, McQuaid CD, Serrao EA, Pearson GA, Jacinto R, Nicastro KR (2016) Upwelling areas as climate change refugia for the distribution and genetic diversity of a marine macroalga. Journal of Biogeography 10.1111/jbi.12744

3b. Likewise, there are other papers for the Iberian Peninsula that have studied long-term variations of species at their range edges, not just for the warm edges in the south but also for the warm edges in northern Spain, where surface seawater warms towards East.

Other suggestions:

4. The index of developmental instability is interesting but can be highly affected by grazing rather than climate stress.  Fucus guiryi are extremely consumed by grazers, namely Sarpa salpa fishes that get to it at very high tides. Did this not affect your measurements? Was this population located so high on the shore that fish could never get to it thus not affecting thallus development? This is a tradeoff with the physical stress factors that increase further up on the shore.

5. The chlorophyll fluorescence parameters are widely used in the literature and do not need to be described in detail, with equations, in this paper that simply uses them.

6. Disappearance and recovery in the same site, so isolated from the remaining populations, suggests that the population might have kept microscopic latent stages (embryos of Fucus can remain undeveloped but alive if conditions are limiting) during unfavourable years, that recovered the population afterwards. This hypothesis has not been addressed in the paper.  See for example our discussion of such issues for an iberian kelp species (despite the differences between these in being subtidal and having gametophytes): Assis J, Coelho N, Alberto F, Valero M, Raimondi P, Reed D, Serrao EA (2013) High and distinct range edge genetic diversity despite local bottlenecks. PLOS ONE 8(7): e68646

7. Is fig. 4 really necessary? It can be simply reported in the text.
Experimental design: no more comments (see above)
Validity of the findings: no more comments (see above)
Additional comments: no more comments (see above)


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

Input:
Title: NORTH ATLANTIC OSCILLATION DRIVES THE ANNUAL OCCURRENCE OF AN ISOLATED, PERIPHERAL POPULATION OF THE BROWN SEAWEED FUCUS GUIRYI IN THE WESTERN MEDITERRANEAN SEA
Review round: 1
Reviewer: 3

Output:
Basic reporting: I found the manuscript well structured, the analyses adequate and sound, the conclusions of an environmental control for species occurrence supported by the data. Text in general was also well written and ideas very easy to follow.
Experimental design: No comment
Validity of the findings: I only have one concern (see also a few minor comments below) relating to the authors interpretation of the nature of the marginal Calaburras population. The authors argue throughout the text that F. guiryi is present every year at least during the winter and that some unfavourable years the population does not survive to midsummer. The sole explanation proposed for the recurrent recruitment during the winter is the putative transport of drifting thalli and unattached embryos from neighbouring Gibraltar populations (some 80km away) by the Atlantic Jet (lines 59-61; 396-401; 424-426). Genetic data shows that Fucus (and canopy-forming brown seaweeds in general) are very poor dispersers at km scales (e.g Neiva et al. 2012). There is a much more parsimonious explanation for the recurring appearance of F. guiry at the very same site – the presence of cryptic, hardy microscopic stages that survive unfavourable months before developing again during milder months or after catastrophic events (Carney & Edwards 2006; Creed et al 1996). Local(not immigrated) few-cell zygotes (something like a seed bank) or remnants of thalli (as in many cryptic perennial Cystoseiracea that lose most biomass during the unfavourable winter months) are good candidates, and both avoid the need for a constant supply of propagules from a 80 km distance source. This hypothesis should be at least discussed as an alternative to the immigration scenario.
Additional comments: Minor comments:
line 48: F. guiryi occurs as far south as Dahkla in the Western Sahara (Lourenço et al. 2016, also discusses the importance of upwelling and lower SST in this African range)

line 49: “sometimes a population develops at Punta Calaburras”. If I understand correctly this is not very clear in the sense that the population is present throughout the years at least during some months and not missing some years

line 52: “(sub F. spiralis and F. spiralis var. platycarpus)”. I don’t understand what these refer to – subspecies? To avoid confusion over the focal species it should be removed or clarified. Same in line 62.

line 67: “university of AFM”- in full if not defined earlier

line 73: “presence at Punta Calaburras – add something like “during/until mid-summer” to avoid confusion. Same in line 394 – add “mid-summer” before “occurrence”

line 166: it could be explicit the institutional source of the data (NOAA?) for AO and NAO, as for SST, instead of just the link

line 423: “persistence until midsummer” would be clearer than “success of occurrence in midsummer”, since the species is there already during the winters. Same in 445 – “proliferates until
midsummer” may be more accurate.

line 449: In calaburras the tidal range is narrower but also overall lower. Stating something like “the narrower, [and overall lower] tidal range” reinforces the differences discussed immediately after

Figure 1: correct “Westarn” Alboran in legend

Figure 2 and 6: clarify UTC in legend

Figure 3: correct “ornage” in legend

References
Neiva et al. 2012 Drifting fronds and drifting alleles: range dynamics, local dispersal and habitat isolation shape the population structure of the estuarine seaweed Fucus ceranoides.

Carney & Edwards 2006 Cryptic processes in the sea: a review of delayed development in the microscopic stages of marine macroalgae

Creed et al. 1996 The development of size structure in a young Fucus serratus population

Lourenço et al. 2016 Upwelling areas as climate change refugia for the distribution and genetic diversity of a marine macroalga


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: NORTH ATLANTIC OSCILLATION DRIVES THE ANNUAL OCCURRENCE OF AN ISOLATED, PERIPHERAL POPULATION OF THE BROWN SEAWEED FUCUS GUIRYI IN THE WESTERN MEDITERRANEAN SEA
Review round: 2
Reviewer: 1

Output:
Basic reporting: This revised manuscript is much clearer to read than the original version -- thank you! However, there are still grammatical English errors throughout the manuscript that need to be fixed. I highlight some, but not all, in this review.  (e.g. the third line of the Abstract reads 'The presence of the alga at Punta Calaburras is supposed could be due to the growth of....' )

The authors should remove the last part of the last sentence in the Abstract 'an aspect practically unknown before'.

Lines 56-57 should be rephrased - unclear.
Line 67: remove 'uninterrupted'
Line 70: remove 'must' -- this has not been experimentally confirmed.
 Remove the two sentences that start on lines 136 and 138 - they are not needed in this manuscript.
Line 143: fix the ...
Lines 172-174: remove these first two sentences, as they do not belong in the Methods section.
Line 184: change 'image' to 'images'
Line 232: change 'cultured' to 'placed'
Line 363: replace 'last' with 'recent'
Experimental design: The authors have not yet clarified how the sites were surveyed for the presence/absence of the algae. Was the entire site visually surveyed? Were quadrats/transects used? The presence of small macroscopic stages may require careful surveying techniques, and this is not clear from the text written.

The authors need to describe how the thalli were selected for the developmental instability analysis. Was this random? Haphazard? Both Random and Haphazard selections of individuals have specific ecological criteria.  The authors need to demonstrate how bias was avoided when thalli were selected.   Similar comment for line 230 in the fluoresence Methods section.

The authors should change dry weight to dry mass, as they were recording mass, not weight.  Also, the equation in line 243 is wrong. The denominator should be fresh mass, not dry mass.  Please note that this equation is no longer Equation 5.   Similarly, Equation 7 is not equation 7 anymore.
Validity of the findings: Line 278-279: rephrase this sentence. A trend of what through time?

Binary logistic regressions (line 280-294) are not linear. I am not sure where the equation came from in line 285, but it does not represent the model output that is shown in Figure 3. This section needs to be rewritten to reflect this.

The authors state that there was not a difference in developmental instability between the two sites, but they had a p=0.04 value for difference by location.  This is significant (as the authors do not state that they used a different p value as a cutoff for significance), but the authors treat it as if it were not. They also do not state which site had the higher developmental instability values.

Figure 2 should also show the tidal range for Pt. Calaburras.
Additional comments: The first paragraph of the Discussion is repetitive from the Introduction. It should be shortened substantially to focus on how the new results from this work fit with current ecological theory on peripheral populations. Additionally, the second part of this paragraph (~371-401) should be significantly shortened for clarity. The authors should simply discuss that the population most likely re-emerges from existing microscopic stages already present at Punta Calaburras, and that dispersal from the Strait of Gibraltar is possible, albeit unlikely.

Remove 434-437 - these sentences are not relevant for this manuscript.

Change the Discussion text regarding the developmental instability results.


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

Input:
Title: NORTH ATLANTIC OSCILLATION DRIVES THE ANNUAL OCCURRENCE OF AN ISOLATED, PERIPHERAL POPULATION OF THE BROWN SEAWEED FUCUS GUIRYI IN THE WESTERN MEDITERRANEAN SEA
Review round: 2
Reviewer: 2

Output:
Basic reporting: The authors have addressed the reviewers concerns adequately.
Experimental design: The authors have addressed the reviewers concerns adequately.
Validity of the findings: The authors have addressed the reviewers concerns adequately.
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: NORTH ATLANTIC OSCILLATION DRIVES THE ANNUAL OCCURRENCE OF AN ISOLATED, PERIPHERAL POPULATION OF THE BROWN SEAWEED FUCUS GUIRYI IN THE WESTERN MEDITERRANEAN SEA
Review round: 2
Reviewer: 3

Output:
Basic reporting: This is the 1st revision of the manuscript by Melero-Jiménez et al. investigating the climatic and oceanographic correlates of mid-summer persistence of a range-edge population of the seaweed F. guiryi and its comparison (developmental and physiological traits) with a less marginal population.

Globally I found the revision adequate as most comments raised were addressed. I only have a few additional minor comments/suggestions that the authors can address without the need for an extra round of revision.

Some ambiguity remains in some sentences concerning the seasonal nature of F. guiryi at Calaburras – it should be clear to the reader that macroscopic F. guiryi is always detected in the winter and either these individuals persist until mid-summer or they disappear, depending on the years. Specifically, that midsummer occurrence is related to survival of winter alga and not recruitment (this happens all years during winter). The authors can consider some suggestions (see minor comments below) that hopefully improve clarity.
Experimental design: R1 mentioned in the first review that it was not clear how presence/absence had been assessed (transects, quadrats, searching haphazardly, other?). This information may be relevant for the readers and should be made available.
Validity of the findings: My personal view (based on available literature) is that migration from Gibraltar is much less plausible than local recruitment. In any case, long-distance (non-local) dispersal by kelp and fucoid seaweeds is much more often mediated by drifting, reproductive thalli (that release their fertilized zygotes in situ after being washed ashore) than by direct travel of spores or zygotes, that in the case of Fucus are actually negatively buoyant (i.e.  they sink). This alternative vector of long-distance dispersal (drifting reproductively thalli) from Gibraltar may be worth mentioning.
Additional comments: Minor comments/suggestions
line 24 – rephrase and correct “is supposed could be due to the growth of resilent, microscopic stages as well as influence of the permanent Atlantic jet”

line 43 – the first sequence is too long; it can be break in two sentences or at least use a coma. It may also be more correct to add “being more abundant (towards warmer range edges) in areas characterized by upwelling”

line 47 – the “known” southern limit would be more correct

line 51, 61- the presence of F. guiryi (named as F. spiralis and F. spiralis var. platycarpus; see Zardi et al., 2011). F. guiryi (named as F. spiralis; see Zardi et al., 2011 was found in 1987…
It may be clearer as “previously named” or “previously/originally identified as” F. spiralis etc…

lines 56 – upwelling causes lower SST and allows persistence in otherwise too hot areas, but this effect does not necessarily have to do with modern climatic change. It may be more correct to rephrase as “in agreement with the hypothesis of upwelling providing thermal refugia/relief for F. guiryi”.

line 67 – “field teaching at the university of AFM”. For clarity it could be added “at the university of the corresponding/senior author AFM”, otherwise readers (like me) may think it is the university abbreviation.

line 70 – Seasonality appears to be exclusive of Punta Calaburras. This could be reinforced as “in contrast to the nearby perennial populations in the Strait of Gibraltar in the Strait of Gibraltar AND the remaining range.

line 87 - replace “population” by “populations”

line 189 – correct “can influences”

line 372-374: this phrase is a bit confusing, please rephrase

line 375: replace “occurrence of thalli” by “re-appearance of thalli”

line 387: “Another, alternative hypothesis is that recurrent re-appearance of F. guiryi at Calaburra is achieved by…” “unfavourable summer months”…

392: local microscopic stages (recruits, holdfast remnants) at PC ca work as a seed bank

393: With our data, we cannot discern whether the recurrent re-appearance of the alga during the winter is due to migration or local recruitment, or both. Taking into…

401: correct F. guiyi

404: “involved in the annual occurrence of the alga” by “involved in the persistence of macroscopic thalli during the summer”

line 421: replace “occurrence” by “survival”

line 461: replace “modulate the occurrence of the annual survival of” by “modulate the mid-summer survival of macroscopic thalli of”

line 470: Dra Ester Serrão


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: INTRASPECIFIC VARIATION IN THE DIET OF THE MEXICAN GARTER SNAKE THAMNOPHIS EQUES
Review round: 1
Reviewer: 1

Output:
Basic reporting: This is a beautiful data set that covers many years.  Well done. The statistical relevance needs to be included in the legends of each figure.  I would suggest attempting to combine the final figure of log snake length and prey mass into one chart.  the use of different symbols or colors would be appropriate, however, this may prove to be too messy and unreadable.
Experimental design: I applaud this manuscript as there are not enough natural history style manuscripts being written.  My sole comment in this section is why were the hemipenes or cloaca not everted to determine sex?  Simply eyeballing the tail breadth will invariably lead to misidentifying the sex of some animals.
Validity of the findings: I would be careful on commenting on the commonness of some of your findings.  You have stated your conclusions well and described the diet of these populations clearly.  I have added some questions in the text, such as what is the percentage of snakes that had food?  I also encourage you to be wary of digestion level of food.  I believe that this has led to many circles on the zero line of figure 3.  I would consider adding this information to the legend or text, or both.
Additional comments: This is a robust data set that you have done a nice job with.  I have identified some concerns in the text and in the notes.  One thing to add is to include a citation for your delineations of snake size classifications.  Have those size classifications been used before, are you classifying them as newborn, etc.. upon personal observation, natural history notes, etc...


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: INTRASPECIFIC VARIATION IN THE DIET OF THE MEXICAN GARTER SNAKE THAMNOPHIS EQUES
Review round: 1
Reviewer: 2

Output:
Basic reporting: The authors have presented a valuable contribution on the diet of T. eques, a relatively little studied gartersnake. They attempt to describe diet differences between sexes and among size classes, which is an important aspect of the species ecology.

This MS needs to have major editing for grammar and language. There are portions that are difficult to follow because of grammar issues.

Authors need to follow consistent format for the species name. As is they switch between scientific and common name.

Authors need to follow proper and consistent format for presenting stats.

I made edits, comments, and suggestions to the PDF. These address questions that need to be clarified and statements that need more justification.
Experimental design: The overall design is simple and appropriate.

I made questions and comments on the statistical approaches that I would like to see addressed.

In the introduction that authors state they pooling older data (Lines 62-64) in their analyses, but it is unclear if this was actually done. As the methods and results currently read the authors only deal with their data. I would like to see them conduct tests with their data, that of the other authors, and the pooled data. Alternatively, instead of doing this independently, they can do a multivariate test using the data source as a variable to determine if diet varied among the data sources. Either way this issue needs to be addressed and clarified.

The authors need to clarify the sex body size analyses because it is unclear to me if this analysis was done with only adults or included all snakes (juveniles and adults). Including all snakes may bias their results to no sexual size dimorphism.
Validity of the findings: As currently presented the results are novel by contributing to our knowledge of the snakes diet between age classes, but I am not completely convinced about the sex differences. See above comment and those to the MS.

I made additional comments to the MS regarding the methods.

Many of the questions and comments made to the discussion may be addressed if other sections are more clearly explained. I found it difficult to connect methods, results, and discussion because of the lack of detail in the methods. Sometimes it is hard to follow the logic and methods the authors use, making further interpretations difficult.
Additional comments: This MS has a lot of potential and the authors have presented valuable data. But in its current state I feel there is more work to do, especially in regards to more clearly explaining the methods and results.

For ontogenetic diet and prey availability discussion I suggest the authors look into the rich body of literature on other Thamnophis 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: INTRASPECIFIC VARIATION IN THE DIET OF THE MEXICAN GARTER SNAKE THAMNOPHIS EQUES
Review round: 2
Reviewer: 1

Output:
Basic reporting: There has been considerable improvement in many areas of the MS.  However, it reads more disjunct with this improvements.  Certain sections suffer from poor grammar while other sections flow very well.  I found it difficult to jump from common name to scientific name throughout the MS.  Please identify the scientific and stick with it.  Line 71, the citations listed are not complete.  i would suggest adding the phrase 'amongst others'.  In addition, Line 73 the author identifies that recent studies do not investigate sexual diff in diet.  i find this to be4 inaccurate and deserves a more comprehensive literature search to address this.  The final paragraph of the introduction is entirely out of place.
Experimental design: The question is appropriate and approached correctly. Line 82: snakes were collected haphazardly at 3 different locales on the Mexican plateau.  the reader does not need to know about flipping rocks, etc..  Line 92:  16 years on 1.5 +- 0.9 occasions each, what is being referred to here?  Line 101 13% of these findings, again, what is being referred to here?
Validity of the findings: The additional analysis performed, the MCA contributes to the robustness of the data.  It demonstrates the data can be analyzed in multiple ways to produce the same results.  I find the MCA analysis much more informative than the cluster analysis and would recommend using that in supplemental materials.  The analysis specifically describes slugs and axolotls on the figures, however, table 1 simply classifies these as others.  It would be best to stay congruent with these data.  Line 233-237 grammar is not present.Line 221 citations are needed because this has been found in numerous other snake species.
Additional comments: I found your improvements and grammar and additional analysis evident.  However, the MS suffers from being very disjunct at this point, to the reader.  I feel like some sections are beautifully written and others are grammatically lacking.  I would suggest a couple of more citations (as noted above).  I think the new analysis adds to the robustness of the data.  I prefer it greatly to the cluster.  Please include examples of the "other" section of table 1 as you address slugs and axolotl specifically in Fig 1.  overall good improvements.


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: INTRASPECIFIC VARIATION IN THE DIET OF THE MEXICAN GARTER SNAKE THAMNOPHIS EQUES
Review round: 2
Reviewer: 2

Output:
Basic reporting: I am pleased with the authors revisions, especially with how the addressed the statistical issues. The authors have done a great job on this manuscript. I made a couple of very minor edits which are noted on the attached PDF.
Experimental design: Addressed concerns.
Validity of the findings: Addressed concerns.
Additional comments: Good job!


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

Input:
Title: WHOLE GENOME SEQUENCING OF RHODOTORULA MUCILAGINOSA ISOLATED FROM THE CHEWING STICK (DISTEMONANTHUS BENTHAMIANUS): INSIGHTS INTO RHODOTORULA PHYLOGENY, MITOGENOME DYNAMICS AND CAROTENOID BIOSYNTHESIS
Review round: 1
Reviewer: 1

Output:
Basic reporting: The text is well written and the article is consistently written in professional english. A  few long sentences may be reframed to simplify the text for readers. Raw data is shared as required and results are well supported with evidences
Experimental design: The methods used are sufficient for the article and the mitochondrial genome elucidation through 'genome skimming' is very innovative. However, I think the readers would like a more detailed account on 'recovery of the complete mitochondrial genome of R. mucilaginosa using the genome skimming approach' as there are few reports on this this and also this is the main attraction of this review. Thee should be more references on similar genome skimming methods used or on other methods for mitochondrial genome recovery. Otherwise, there are several reports on description of the genome and transcriptome of Rhodotorula.

I didnot follow why the authors have focussed on Distemonanthus benthamianus in detail. I think the details should be minimized as the site of isolation has no effect on the description of genome. If there is some significance of this plant in this context then it should be clarified in the article
Validity of the findings: Data is robust and although there is nothing much to be 'newly' reported, the mitochondrial genome recovery does give this article an edge above the other articles/reports published on genome of Rhodotorula.
Additional comments: The results and discussion section and conclusion section can be made more informative by
(1) comparison to other similar reports
(2) more description on genome skimming
(3) comparing the presence of carotenoid pathway gene(s) in yeast Rhodotorula and plants or bacteria, since the article is more focused on carotenoids production and its importance as a valuable anti-oxidant
(4) I believe readers would also want to know details on "homing endonuclease from the LAGLIDADG and GIY-YIG families" and a section can be added on this


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: WHOLE GENOME SEQUENCING OF RHODOTORULA MUCILAGINOSA ISOLATED FROM THE CHEWING STICK (DISTEMONANTHUS BENTHAMIANUS): INSIGHTS INTO RHODOTORULA PHYLOGENY, MITOGENOME DYNAMICS AND CAROTENOID BIOSYNTHESIS
Review round: 1
Reviewer: 2

Output:
Basic reporting: The manuscript describes the isolation of a putative endophyte from the plant Distemonanthus benthamianus, its identified as a strain of Rhodotorula mucilaginosa, genome sequencing and analysis of the genome.  Primary observations from the genome were the annotation of the mitochondrial genome and identifying the candidate genes for synthesis of carotenoid molecules.

The manuscript is a solid set of data.  The following points are aimed to improve the text.

A general point is that the preexisting literature has been someone sparsely covered in places, and that then limits some of the conclusions that can be drawn.  For example:

(1)	Line 58 species within the genus and “validating the monophyly of the genus Rhodotorula” and line 307.  Here consideration of the two Wang et al. 2015 papers in Stud Mycol. 81: 27-53 and 149-89 would be wise, since these were the study to clean up Rhodotorula, as until then the genus was polyphyletic.  The conclusion on line 307 is too strong without all the species in the genus being included.
(2)	Lines 94-97: a key point is that the genes have been mutated and shown to be involved in synthesis of pigment.  This was first done in the closely related genus Sporobolomyces (Abbott et al. 2013 Appl Microbiol Biotechnol (2013) 97: 283-295), and then the two genes in Rhodosporidium (=Rhodotorula) toruloides (Koh et al. 2014 BMC Microbiol 14:50; Sun et al. 2017 Biotechnol Lett 39: 1001-1007).
(3)	Strain RIT389 should be deposited in one or more culture collections, e.g. CBS.

Text edits:

Line 83: write as “resources for R. mucilaginosa are surprisingly scare in public databases”.
Lines 83-84: JGI has also sequenced R. mucilaginosa strain ATCC58901.  Given the discussion on the genus, it may be worth pointing out that there are genome projects for other Rhodotorula species.
Line 91: “in at least three different fungal species” is too vague.
Lines 107-108: Replace “A relatively recent study showed that” with “The”.
Lines 192-193: how the identification was may should be clearer, i.e. PCR of ITS and sequencing.
Lines 197-198: there appear to be two unrelated points in this sentence, so better split into two.
Line 213: add “levels” for “carotenoid levels when”.
Line 226: “i.e.” in place of “e.g.”.
Lines 233, 235 and 239: spelling of “toruloides”.
Lines 237 and 241: using “Rhodotorula sp.” would be better.
Line 274: “Identification of a genomic region”.
Lines 280 and 281: better with the addition of two commas, as synthase, and carotenoid, .
Line 304: “databases” plural.
References: check for formatting, e.g. italics on species names, remove capital letters in article titles.
Experimental design: Lines 187-192: (a) This section needs more background on the isolation of the microbe from the chewing stick.  (b) While the description may have been what happened, it does not present the authors in a strong light.  SEM is over-kill to show something is not a bacterium.
Validity of the findings: See comment 1 in section 1 about the claims on monophyly of the genus Rhodotorula.
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: SNAPSHOT RECORDINGS PROVIDE A FIRST DESCRIPTION OF THE ACOUSTIC SIGNATURES OF DEEPER HABITATS ADJACENT TO CORAL REEFS OF MOOREA
Review round: 1
Reviewer: 1

Output:
Basic reporting: The article is clearly written and provides appropriate references. The figures make sense, although I would like to have supplemental material with a few acoustic recordings so that we could listen to the different soundscapes.
Experimental design: I find 2 major flaws with the design which limit the conclusions one can draw from the study:

1) Temporal window: I am concerned that recordings were only made during daytime hours. I think this is a big oversight. You mention that there is high variability during nighttime and early morning hours – but this is exactly when you may see larger differences between habitats. For example, if a species of soniferous fish lives only in deeper water, but they only sign at night, then you would not pick up this difference between depths/habitats by missing their prime singing window.

2) Temporal replication: I understand that perhaps logistical or financial constraints were at play here, and the investigators may not have had access to longer-term recorders or the ability to dive in deeper waters. But 10-minute recordings during subsequent mornings is not very high replication. I think the authors should emphasize the “snapshot” nature of these recordings and put this caveat in their results and possibly in the title.

That said, there is still value in this work, but it doesn't knock your socks off in terms of novelty or  a grand contribution to the field.

There are also a few points of clarification needed:
-exact water depth of recordings (in a table)
-why no inclusion of higher frequency sounds
-how the spectral averaging was done
-did you look/listen to recordings and get any explanation for some of these differences/sound sources?

One other point  about experimental design/analysis:
 Did you try using any of the newer eco-acoustic indices like acoustic complexity, acoustic dissimilarity, etc? I am guessing that you may have tried, but found no significant patterns or perhaps even misleading results. You would not be alone – several others have tried and have learned that these indices are not “ready” for the marine environment yet. There was a lively discussion about this at a recent ASA meeting, and a recent paper by Staaterman et al (2017, MEPS) demonstrates an example in which the indices don’t work as expected. I know many authors are reluctant to put null results or confusing results into a manuscript, but I would urge you to include these analyses anyway. It would bring further emphasis to the fact that these indices require further ground-truthing for the marine environment.
Validity of the findings: Some of the interpretation of the differences between habitat types and distances from shore need to be refined. There needs to be a discussion about the propagation of low-frequency sounds in shallow water.

Otherwise, the conclusions are well-stated and reasonable.
Additional comments: Given the fact that the authors probably cannot go back and redo these recordings, or perhaps were limited with their instrumentation, it's not reasonable to ask them to increase replication. This is a small study but still a valid contribution to the field. For this reason I am putting "minor revisions" rather than "major revisions". But I would like to see them try running the newer eco-acoustic indices (with existing data), and to clarify about some of their methods for spectral averaging etc. And I encourage them to think about higher spatio-temporal resolution in future work, if time and money allows.

small comments here:

Introduction – line 48: you start two sentences in a row with “as an example”

Methods: - line 93- is Tiahura a natural canal or man-made? Why are there no corals there?

Line 110: I am concerned that recordings were only made during daytime hours. I think this is a big oversight. You mention that there is high variability during nighttime and early morning hours – but this is exactly when you may see larger differences between habitats. For example, if a species of soniferous fish lives only in deeper water, but they only sign at night, then you would not pick up this difference between depths/habitats by missing their prime singing window.

Please give more specifics about the water depths where recordings were made. I see that you have the range of water depth for each habitat type, but this range is pretty big (~20m).

Methods: so the boat was drifting, not anchored? To me this is another potential source of error. I have made recordings of the hull of a boat and the slapping/lapping of waves is indeed a problem. Were you not able to deploy passive recorders for longer periods?? How did you deal with these lapping noises?

Why did you not include analyzes of higher frequencies (snapping shrimp band)? I understand perhaps more interest in looking at fish vocalizations, but it would not be hard to also include snapping shrimp here.

I understand that perhaps logistical or financial constraints were at play here, and the investigators may not have had access to longer-term recorders or the ability to dive in deeper waters. But 10-minute recordings during subsequent mornings is not very high replication. I think the authors should deemphasize the “snapshot” nature of these recordings and put this caveat in their results and possibly in the title.

Lines 143-152: please clarify language here. It is not clear how you got the 12 samples. Also, please explain the “averaged for each sampled frequency” – did you not have power spectral density in 1-Hz bands? From the way your axes on the plots are labeled, it appears that you did not have 1-Hz bins and some averaging across frequencies took place. Please explain what bandwidths were used. Does the recorder automatically spit out SPLs and not the raw waveforms?  This part needs clarification.

Line 163: I find the phrase “distant habitat” misleading here. You mean distance from shore, I assume, but you may also mean distance from other habitats. I would instead say “deeper habitats”. If you don’t mean distance from shore, this would imply that your hydrophones were not truly “in” the environment you claim to be sampling.

Lines 222-234: I think some discussion is required about the propagation of low-frequency sounds in very shallow water. Since transmission of these sounds is restricted by water depth, you would actually expect to have lower sound levels in these lower frequencies for the shallowest sites. The fact that you got the opposite demonstrates that there is a greater contribution from geophony or biophony.

Line 235-237: I don’t see how your results lead you to this conclusion. Higher sound levels at different habitats along transect 1 could mean more sound-sources at the other (non-reef) habitats, which you state later in the paragraph. I would remove this sentence or rephrase it. I would also mention later in this paragraph the fact that deep water enables transmission of lower-frequency sounds, so in addition to the fact that there may be additional sound sources in the reef slope/sandy area (fish), it may also have to do with the physics of the environment.

Did you try using any of the newer eco-acoustic indices like acoustic complexity, acoustic dissimilarity, etc? I am guessing that you may have tried, but found no significant patterns or perhaps even misleading results. You would not be alone – several others have tried and have learned that these indices are not “ready” for the marine environment yet. There was a lively discussion about this at a recent ASA meeting, and a recent paper by Staaterman et al (2017, MEPS) demonstrates an example in which the indices don’t work as expected. I know many authors are reluctant to put null results or confusing results into a manuscript, but I would urge you to include these analyses anyway. It would bring further emphasis to the fact that these indices require further ground-truthing for the marine environment.

A note for future work: I have been to Moorea and noticed the big cruise ships that come through. I would be curious to know how the propagation of the ship noise differs depending on geographic location, again due to the physical constraints of shallow vs. deep water. I would suggest you put out some passive recorders at different depths/transects along the reef slope and see what happens when a ship comes through. Would also be interesting to put some inside those two bays/cracks in the island.

Figure 5: could be displayed better – consider using color and putting all on one plot, or adding another panel with average for each habitat type.

Would be nice to have supplemental data with recordings from each habitat type.


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: SNAPSHOT RECORDINGS PROVIDE A FIRST DESCRIPTION OF THE ACOUSTIC SIGNATURES OF DEEPER HABITATS ADJACENT TO CORAL REEFS OF MOOREA
Review round: 1
Reviewer: 2

Output:
Basic reporting: Intro and background are sufficient to outline the problem. In general, the manuscript would benefit from review from a native English speaker for improved clarity and word use. A summary table of the raw data was supplied (although the raw data itself, i.e., wave files) was not. I am not sure if this conforms to PeerJ’s standards. Figures 3-4 are non-normative and I am not sure what they add in terms of interpretation.
Experimental design: I have three major problems with this study. Data collection during the day is obviously totally insufficient because this is not, as the authors note, the time when most soniferous organisms on reefs are active. Furthermore, it seems from the methods as if the transects were conducted sequentially and within transects the stations were visited south to north. Then this was replicated in the same way two more times. If that is true then there is a major bias there in terms of the times at which each station and transect were sampled. Finally, the authors attempt to compare the recordings they made to habitat type but bring no evidence of the fauna resident in each habitat type. For these three reasons, I cannot recommend publication of this research.
Validity of the findings: For the reasons stated above, I do not interpret these findings as valid.
Additional comments: Lines 82-83: “highly specialized habitat requirements” – such as?
Lines 82-85: the logical link between “highly specialized habitat requirements” and variations in low-frequency SPL is not clear. Suggest rephrasing.
Line 94: “noisy” – first of all this is somewhat subjective, but 90 dB is not, in my opinion, “noisy”, or, more appropriately, high amplitude
Lines 98-105: could the authors please bring evidence to support their claims about the species richness of each of these habitat types. Did they conduct visual surveys? Or is this conjecture?
Lines 131-133: The close to shore station was always visited first in each transect and replicate? If sound levels change throughout the day then this would introduce a major bias into these results. Did the authors not put recorders at each station for the entire day or ideally over several diel periods? If not, there is no way to know how sound levels were changing during the sampling period.
Results section: I found this entire section very hard to understand.
Lines 231-234: True, it is very important to consider the physical environment and its influence on ambient sound. Because the authors do not have visual survey data to link to fauna at each reef, it seems as if purely physical differences among the stations would be the most parsimonious explanation of the differences found.
Lines 258-262: I agree with the statement here that the short sampling period is insufficient to properly characterize these habitats. A large body of literature now exists that points to the importance of diel sampling because of how dramatically sound levels can change even over short temporal scales.


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: SNAPSHOT RECORDINGS PROVIDE A FIRST DESCRIPTION OF THE ACOUSTIC SIGNATURES OF DEEPER HABITATS ADJACENT TO CORAL REEFS OF MOOREA
Review round: 1
Reviewer: 3

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The Ms provides a first insight into the acoustic features of different habitats adjacent to the coral reef surrounding the north coast of Moorea (French Polynesia). In particular, it focuses on the description of soundscape signatures (sound energy along frequencies) along three transects of four recorded points each (barrier reef; sandy plain; reef slope; drop-off).
The Ms is well organized and the results produce interesting advance to our knowledge of the field. These baseline data are strongly needed to produce a benchmark that could be used as a reference for future biomonitoring studies. Here I provide a list of recommendations to improve the MS.

The authors have provided a good temporal and spatial diversification of samples by investigating three points for every habitat, and replicating in three different days. Anyway, the total amount of recorded minutes in each of them is quite limited. Soundscape holds a great deal of variation over time of the day and over days. It could substantially change either to the time of the day in which recordings were taken (for instance many fishes vocalize in specific temporal windows) or may also vary from one day to the next (see Pieretti et al. 2016). In order to capture a reliable example of the soundscape of the area, longer recordings would be the optimum solution.
The authors’ dataset is limited to 12minutes (divided in 4mins in three replicates) for every recording station. That makes 36 minutes for every habitat type. Moreover, authors do not fully specify the recording time of the day (line 110: Recordings were conducted between 09:00 and 16:00).
I still consider the collected data a useful and needed starting reference for future studies, however in the discussion I suggest to explain this limitation (adding details at line 261) or to the reasons that have led authors to consider it as less important  (maybe in these tropical environments soundscape fluctuations are weak).

I recommend that a native English-speaking colleague review the Ms. In fact, examples of some errors and typos along the paper are at line 93 (depths), 110 (recordings), 181 (did not differed) or the sentence at line 101-103 should be rephrased.

The statistics should be better presented and clearly explained. At lines 153-160, the current paragraph makes its understanding challenging. I suggest specifying the number of cases (N) - embedded in the text or in the caption of figures – so that the reader can easily understand which data have been considered for each analysis.

Others:
Keywords are missing.
Line 143: FFT 1024 on the 48kHz? (line 130) Figures seem to have a higher resolution in frequencies.
Line 143-144: I would explain why this step was taken
Line 173: I would refer to figures before
Line 179: from
Lines 224-226: it was not stated earlier
Figure 1: rescale the distance among transects; the distance among 2 and 3 seems to double the distance of transect 1 and 2
Figure 5: Caption: refer to the statistics used

Dr. Nadia Pieretti
Dipartimento Scienze Pure e Applicate - DiSPeA
Università degli Studi di Urbino "Carlo Bo"
Campus Scientifico Enrico Mattei
Località Crocicchia
61029 Urbino
Italy


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: COLOUR CHANGE OF TWIG-MIMICKING PEPPERED MOTH LARVAE IS A CONTINUOUS REACTION NORM THAT INCREASES CAMOUFLAGE AGAINST AVIAN PREDATORS
Review round: 1
Reviewer: 1

Output:
Basic reporting: The paper is well written and covers the relevant literature.
Experimental design: The experimental design is suitable, though as mentioned below, the authors should justify the switch from spectroscopy to objective imaging (without animal-vision) for the heterogeneity experiment.

The authors use the term “chroma” interchangeably with hue without justifying why. In any case, this should be changed or clarified, and will not affect the overall interpretation.
Validity of the findings: The findings are valid, though there are some areas that require clarification, or could be improved:

A tweak in the “greenness” calculation used might improve the findings and reduce statistical dispersion issues (see general comments).

I think the authors should be much more careful when asserting that the larvae were not adopting an intermediate colour in the heterogeneous treatments as they are unable to prove that the background is heterogeneous from the larval perspective instead of plausible mechanistic or behavioural alternatives.

Statistics: using ordinal statistics might offer a more clean way to analyse the result of different treatment levels (e.g. cumulative link models using the ‘ordinal’ package in R allows for mixed models), though I think the author’s approach is ok. As mentioned above though, the ‘greenness’ measure used creates a large skew in the dataset, which means the green treatment will have a disproportionally more powerful influence on the fitted models than the other treatments. In practice the author’s findings are very clear in that there are intermediate levels of greenness and luminance on the different treatments.
Additional comments: Why was photography used for the heterogeneous treatments, while spectrometry was used for all other treatments? This needs justifying. The images were not converted to bluetit cone-catch quanta images, so will be different to the spectrometer-based measures and the authors should make this more clear to the reader.

The use of imaging means the authors could also look at within-individual heterogeneity. e.g. rather than measure average larval colour, they could see whether larvae on heterogeneous backgrounds also have greater variation. To do this the authors could divide the G channel by the sum of RGB channels, then measure the standard deviation in pixel levels. Higher standard deviations would indicate higher within-individual variation in green and brown patches. This extra analysis would add some useful extra information, though is not essential.

Habitat specialisation in the heterogeneous environments provides a plausible explanation for the finding that larvae tended to specialise on one background colour over another. e.g. the larvae wouldn’t be expected to switch to an intermediate colour if they could always choose to sit on a single background type. Given the authors were unable to track individual movements I think they should present this as the most likely strategy. If the dowelling were striped (so that the larva’s bodies are always crossing many colour patches) the intermediate strategy might be more believable, however even then it could be due to a simple mechanistic constraint that only allows the larvae to sense very small patches of their substrate colour near their eyes/sensing organs.

L152-163 The definitions of colour and luminance used here are mentioned before talking about the visual system being used, and how luminance is calculated (i.e. bluetit double cone catch quanta under a given illuminant). Chroma would normally be the distance from the centre of a triangular (trichromatic) or tetrahedral (tetrachromatic) colour space, and hue would be the angle(s), but the authors seem to use chroma to refer to both hue and chroma (which is often called saturation). I think the authors should change their definitions to more standard ones, or justify why they are combining hue with chroma. To make matters more complicated chroma is later defined as ‘greenness’ in L176.

L157-158 What exactly is being being measured in these statistics? Green and brown dowels would probably differ much more in hue more than chroma.

L210 This calculation of a ‘greenness’ opponent channel has been used by others before, though using MW/(MW+LW) would generate numbers that are more biologically relevant (symmetrically bounded between zero and one, instead of asymmetric bounding between zero and infinity). The calculation of greenness from the photographs later on does this correctly. This might, for example, result in a better fit in figures 5 and S2, which shows a disproportionate effect by the most green treatment.

L216-221 How were the images linearised (the authors should cite the paper and methods used to do the objective image analysis)?

Figure 3 I think the y-axis should be labelled “larvae greenness” if this is correct. Also, the dowel measurements should be labelled and perhaps segregated more clearly from the larvae measurements.

L240-241 and 254-256 The use of an asymmetric greenness measure (as mentioned above) means that these discrepancy values cannot be compared and should probably be replaced with something more meaningful. i.e. the greenness measure is certainly not a perceptually uniform colour-space in which to look at distances. Distances in tetrahedral colour space are also somewhat problematic as these too are not perceptually uniform. Calculating JNDs would be the easiest way to quote colour differences (and would probably be useful as it is biologically meaningful).


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: COLOUR CHANGE OF TWIG-MIMICKING PEPPERED MOTH LARVAE IS A CONTINUOUS REACTION NORM THAT INCREASES CAMOUFLAGE AGAINST AVIAN PREDATORS
Review round: 1
Reviewer: 2

Output:
Basic reporting: .
Experimental design: .
Validity of the findings: .
Additional comments: Colour change of twig-mimicking peppered moth caterpillars REVIEW

GENERAL
I am happy with this very interesting MS that has been thought out conceptually and tested carefully. Excellent literature review in Intro but seems rather duplicated in Discussion – you need only say it once, probably most of it in Discussion.

I would like a slight change of emphasis in the Introduction to explain why it is so important to explore the issue of slow colour change. It makes an attempt at this but gets lost somewhere in the prose. You may want to cut the Intro length while doing this – it is really very long at present – aim for 3 paras. For example only last 2 sentences of first para really needed! Paras 2 and 3 are really about mechanism – not what is covered in this MS.

There is remarkably little said about masquerade in the MS which is odd – after all, these organisms resemble twigs to our eyes anyway. Would we expect greater or less background matching in masquerading than non-masquerading species? Is it a synergistic trait, or a trade-off between shape and colour. Aren’t there comparable data with Kallima butterflies and striped skunks – and probably more that I don’t know of? You might want to discuss these alternatives ways of looking at the colour/shape masquerade relationship in the Intro?

It would be good to have more natural history: are these larvae sedentary on one plant – are there different parts of the plant with different colours? What are the challenges faced by individual larvae in terms of colouration?

Are there any data on this model species that pertain to larval colour and the industrial pollution story?

Figure 1 excellent

SPECIFIC
23 larval stages unclear
35 rigorously is a bit strong
168 what is a B&Q?
291 How do you know visual capabilities?
304 ref not in italics
360 any anecdotal evidence for weak selection?
Table 1 to SOM

Tim  Caro


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: COLOUR CHANGE OF TWIG-MIMICKING PEPPERED MOTH LARVAE IS A CONTINUOUS REACTION NORM THAT INCREASES CAMOUFLAGE AGAINST AVIAN PREDATORS
Review round: 2
Reviewer: 1

Output:
Basic reporting: basic reporting - pass
Experimental design: experimental design - pass
Validity of the findings: Validity of the findings - pass
Additional comments: The authors have been very thorough in addressing all of my concerns and queries, and I am happy to recommend acceptance. They have done a good job of acknowledging the minor limitations of their study, and making these clear to the reader.


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: COLOUR CHANGE OF TWIG-MIMICKING PEPPERED MOTH LARVAE IS A CONTINUOUS REACTION NORM THAT INCREASES CAMOUFLAGE AGAINST AVIAN PREDATORS
Review round: 2
Reviewer: 2

Output:
Basic reporting: .
Experimental design: .
Validity of the findings: .
Additional comments: I have carefully read the revised manuscript entitled: Colour change of twig-mimicking peppered moth larvae is a continuous reaction norm that increases camouflage against avian predators. The authors have made considerable efforts to address the points raised by the editor and 2 reviewers, and I am happy with the outcome. I am not sure if the Conclusion section is needed but otherwise it is ready for publication.


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

Input:
Title: GENERATION AND CHARACTERIZATION OF CROSS NEUTRALIZING HUMAN MONOCLONAL ANTIBODY AGAINST 4 SEROTYPES OF DENGUE VIRUS WITHOUT ENHANCING ACTIVITY
Review round: 1
Reviewer: 1

Output:
Basic reporting: The report is clear and contains relevant results. However, language could be improved and revision is needed.
Experimental design: The research question is well defined and the results support the conclusion. Some terminology would have to be spelt out.
Validity of the findings: Although the study data is robust, the impact and novelty of the results are not immediately clear. While there are many findings on neutralizing antibodies to DENV for therapeutic usage, the authors would need to highlight the utility and  novel functions of the antibody in this study.
Additional comments: Major
(1)	The manuscript was well written and the experiments are well defined and support the conclusion of the study. While neutralizing antibodies are critical in disease protection, your introduction needs to be further clarified as to why the epitope characterization is needed, as some part of the study has been done by other investigators. The objective would need more clarity, for example, are you performing the experiments to develop a potent neutralizing antibody with no ADE activity or are you trying to determine the antibody epitopes?
(2)	Because the antibody used was developed from a previous study, please clarify which parts of the study (if any) contains data from the prior study.
(3)	It is known that ADE activity hampers neutralizing activity of an antibody. Please explain why both the modified and original antibodies demonstrated similar NT levels, despite the absence of ADE activity in the modified antibody.
(4)	Would the concentration of antibody needed for virus neutralization/disease protection be feasible? How much antibody would be required to neutralize DENV at different MOI?
Minor
(1)	Try not to use abbreviations in the title, it would be better to spell out ADE.
(2)	The manuscript needs to be revised for spelling and grammar. Some examples where language could be improved include lines 239, 246, 292, 319.
(3)	Describe PhD-12 and C7C. Were they the identification codes for phage display?


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: GENERATION AND CHARACTERIZATION OF CROSS NEUTRALIZING HUMAN MONOCLONAL ANTIBODY AGAINST 4 SEROTYPES OF DENGUE VIRUS WITHOUT ENHANCING ACTIVITY
Review round: 1
Reviewer: 2

Output:
Basic reporting: Paper is poorly written, English is not clear and unprofessional. Many grammatical, incomplete sentences, poorly chosen words, and conceptual mistakes (e.g humeral instead of humoral) of the English language are found through all the paper’s length. For example (and only 3 examples):
Lines 108-109 Purified B3B9 HuMAb was used in phage display panning experiment to characterize their binding epitopes
Lines 286-288: After comparing this peptide with the amino acid sequences of E proteins of DENV1–4, it was found that all these LXXXG motif was matched to 107LFGKG111…
Lines 393-394: Moreover, N297Q mouse MAb can competed enhancing antibody in polyvalent serum in in vitro study (Williams et al., 2013).

A proofread procedure by an English native speaker is strictly needed.

The title should include the specificity of the MAb characterized to enhance the relevance of the study. For example: “Characterization of aglycosylated human monoclonal antibody targeting the fusion loop of dengue E protein shows strong neutralizing and no enhancing  activity“

Introduction:
The introduction is lacking important information:
1.	The IgG glycosylation patterns and its interaction with the Fc-gamma receptor is needed in order to expose the importance of the Fc modification in the Mab. Literature from Nimmerjahn F and Ravetch is suggested.
2.	“Dengue disease, transmitted by mosquito-borne infection,…” is poorly described. Dengue virus is transmitted by a mosquito, dengue disease is the result of this infection.
3.	The aim of the study written in the last paragraph is confusing and a stronger statement regarding the title of the manuscript is suggested. Through all the manuscript the main objective of the paper is not clear. Even though there was a lot of work invested and the results are interesting, the paper lacks the proper communication of its message and there is not a clear relevance of its findings.

Figures:
Figure 1c, 2 are not of high quality, they look blurry.
In Figure 2 is the control just PBS instead of the antibody, a proper control is a staining including non-infected cells.
In Figure 4. Please show first the ADE in K562 and in B the THP-1 as it is explained in the text. It may seems that A is B and B is A if the methods and results portion is correct. K562 ADE was measured by counting infected cells and ADE in THP-1 with RNA quantification. Which by the way, is not a quite accurate method for measuring Dengue, since there are a lot of RNA products that are not part of an infectious virus particle.

Hypothesis and scientific question is not clearly expressed and needs to be more accurate.
Experimental design: Even though the paper is interesting, it is a purely an instrumental paper constructing a potential anti-dengue therapeutic antibody, but it is of concern if it is enough for the PeerJ journal. Maybe if the paper would be improved in clearly stating its objective, it can be taken for consideration for publication.
A more profound justification of the reason for choosing the Phage display methodology is needed. Concentrations and virus moi are missing. In Figure 1, an antibody without the specificity in the competititve Elisa is needed as a negative control.
The research question is not well defined. From the results, the reader can infer the biological relevance of the paper but it is not clearly stated.
The exact reference in some cases is missing, for example, on has to infer the  original work showing the origin and description of the Mab B3B9.

Results are written with scarce findings and conclusions. Some controls are lacking. For example:
-       demonstrating the absence of glycosylation in the Ab is needed.
-       showing the non/binding to the Fc gamma receptors by FACS is missing.
-       The specificity of the MAb should be confirmed by an assay showing the interaction between the MAb and the fusion loop.
-       Treatment of the original MAb with enzymes that removes the glycosylations would give more robustness to your results.
Validity of the findings: The potential impact and novelty in this manuscript is not assessed by that authors. Even though there is a potential newly produced therapeutic antibody, the paper lacks to send its message.
The asseverations of having constructed a new therapeutic antibody are ambitious regarding the results that they are showing. No proper controls are shown. The biological relevance of an antibody cannot be limited only to its neutralizing or Fcgamma receptor binding. Therefore, the paper needs to be concluded in a more conservative way.
Additional comments: Even though the paper is not well written, the message is not clear and the biological relevance of this new antibody is purely speculative, if the paper is rewritten in a clearly manner, some of the methods and results are more extensively described, if in the discussion they will be able to suggest the potential of this Ab but lowering their asseverations, the paper could be published.