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: ESTIMATING THE DIFFERENCES IN CRITICAL THERMAL MAXIMUM AND METABOLIC RATE OF HELICOVERPA PUNCTIGERA (WALLENGREN) (LEPIDOPTERA: NOCTUIDAE) ACROSS LIFE STAGES
Review round: 1
Reviewer: 2

Output:
Basic reporting: The manuscript is written fairly clearly and the literature referenced is relevant and adequate. Some changes to the English are needed in places. Article structure is good, although the figures could be improved. The manuscript is self-contained with relevant results and hypotheses. Specific comments are provided in the general comments to authors.
Experimental design: The research questions are well defined and interesting. Measurements are done well and most aspects of the methods, especially the respirometry, are reported with great details. I have concerns about the adequacy or reasons for the low sample sizes and the limitations that this has for the study, which are detailed in specific comments that are provided in the general comments to authors.
Validity of the findings: The findings are clear enough, but are undermined by low sample sizes and low power that reduces the robustness of the findings. Figures and in text results could be more clearly aligned if means were shown together with the boxplots. The data provided demonstrate another finding that mass loss during thermolimit respirometry occurs differently for the different life stages - this in context of water loss and CTmax could be reported and discussed. The limitations of the findings because of low sample sizes, in my opinion, should at least be acknowledged.
Additional comments: This study investigates the upper temperature limits (CTmax) and metabolic rates (VCO2) of different life stages of a pest insect, Helicoverpa punctigera. The authors found differences in the life stages for both CTmax and VCO2, but no differences between males and females. The manuscript addresses questions that I think are important to ask in the thermal tolerance space – differences with sex and life-history, and of pest species. The manuscript was an interesting read and the methods to measure thermolimit respirometry and detect activity are really neat. However, there are many points in the manuscript that I think need clarification or better discussion and there are two significant points that are the sample size and the lack of connection of the CTmax and VCO2 rates with the change in mass during the measurement. I hope that my suggestions and questions are useful for the authors when revising their manuscript.

1)	Sample sizes are very low and diminish the conviction with which these results can be stated. It is fortunate in a way that the results are quite clear despite the low power, but the potential for making false positive errors is reasonably high. For instance, even one or two more individuals could change the statistical significance of the results in some cases (e.g. the life stage x sex interaction with CTmax in mean differences reported in text vs boxplots in the corresponding figure illustrates how tenuous this is). This is a shame because I think there is great potential value in the questions that this study asks and the data that has been gathered.

2)	From the data provided, you can calculate mass loss during thermolimit respirometry, which differs substantially among life stages and is also relatively high (a quick calculation and glance at the numbers shows ranges of mass loss as ~13-21% in adults, ~20-24% in larvae, and ~1-3% in pupae). Although this leans into speculation, one could interpret this in a few ways. E.g., as passive differences due to changes in spiracle function/structure across life stages and as a result of different rates of evaporative water loss due to respiration. Or it could be simply due to the respirometry methods where the very dry (Drierite-scrubbed) air increases water loss potential in the insects. Was water measured by the Li7000 during measurement? However, could it also be an active process of using evaporative water loss to cool down at high temperatures and could therefore be increasing CTmax? This is certainly something that occurs in plants in response to high temperatures is there is sufficient water available to lose and I suspect the larvae stage has the greatest water resource availability (from constant foraging) to potentially employ this strategy. I think the change in mass data could be worth thinking about more, including, and discussing in this context, as it adds potentially useful explanatory information about differences in CTmax and VCO2 across life stages.

Specific points:

L38: the word ‘impacts’ is not needed as ‘impact’ already occurs earlier in the sentence

L59-60: I find this counterintuitive - a slow temperature ramp leading to heat shock – the ‘shock’ is likely to be stronger if the organism is abruptly introduced to a high temperature or rapidly increased to that temperature. The other process that can occur during a slow ramp is heat hardening or short-term physiological adjustments (e.g. induction of heat shock proteins) that protect cells from heat damage and might increase CTmax).

L64-67: but apart from this mention where you say that “thermal performance curves must be used”, you don’t then use or even discuss thermal performance curves again, so doesn’t this undermine your approach? If it is really necessary, then it should have probably been done – whereas I think there is value in CTmax and thermolimit repirometry without it, so consider changing the wording around this to be less self-critical.

L72: although it makes good sense and it may or can increase the range of environments an organism can function or survive in, “plasticity is often highly adaptive” is a commonly held assumption but one that needs further substantiation. For example, thermal plasticity in animals and plants has some but relatively limited support for having direct (positive) effects on fitness (necessary for it to be adaptive in an evolutionary sense) - see Arnold et al 2019 Phil Trans B 10.1098/rstb.2018.0185 for a review on the topic.

L68-79: as with the comment above, this paragraph would benefit from a very clear definition of what is meant here by “physiological plasticity” – it starts with thermal phenotypic plasticity and then goes into physiological plasticity – but are they the same in the context of the paper? Often people refer to plasticity but aren’t specific about what trait or aspect of biology is plastic, and this is the key point: there are many aspects of an organism’s biology that can be plastic, but only some of which will likely matter in context of response to changes in their environment. I think it is worth clarifying your use of terminology and definitions, including physiological plasticity, thermal variation, and fitness to be more specific and appropriate to the context of your study.

L85: change ‘modify’ to ‘to be modified’ and L86: ‘narrow’ to ‘narrowed’

L105-107: phrasing is confusing

L108-110: could combine these two sentences to remove redundancy

L117-121: specific dynamic action should probably be referred to in the first sentence of this paragraph where it is being defined.

L127: not sure about use of the phrase ‘The unique CO2 output’

L142-143: this sentence and reference to work on metabolic rate responses to insecticides doesn’t inform the reader of the results of what happens to metabolic rates with exposure or more important why this is relevant other than being a recent paper from the lab group.

L144-146 is redundant with L154-155.

L148-149: very cool feature that sets the temperature and exposure time relevant to body size and surface area!

L159: might be worth pointing out here or earlier when burrowing is mentioned that the reason for this is that the soil the pupae is burrowed in is far more thermally stable than leaves.

L209-210: 0.25˚C/min * 140min = 35˚C increase, but the ramp was only 25-55˚C (30˚C is the range), so some number here must be off and should be corrected, or if the temperature is held constant at the hottest temperature or something like that, it should be mentioned.

L224-225: only 5, 10, 10 individuals for each life stage is very few data points… From experience I completely understand that measuring VCO2 with flow-through respirometry and using Li7000s can be very slow throughput (~ 1 individual / 2-3 hours), but even so, these sample sizes (especially for a pest insect species that is reared in the lab) are very small and could be problematically so for analyses of the data and their subsequent interpretation, so justification for so few individuals is really needed.

L251-255: please clarify which measurement of body mass was used (before or after measurement are provided in the dataset). As in my major suggestions, I think another analysis of the change in mass from thermolimit respirometry would be useful and simple to include in this study since the data is already available. In a broad sense, there is good reason to include body mass in the models for metabolic rate (allometry: Darveau et al 2002 Nature 10.1038/417166a; White et al 2019 Nat Ecol Evol 10.1038/s41559-019-0839-9). However, I think it is also worth including body mass in the final model of CTmax and reporting the lack of relationship properly rather than saying it wasn’t significant and therefore excluded. Recent studies have investigated the patterns of CTmax and body size across species with some mixed findings that could be worth discussing (e.g. Leiva et al 2019 Phil Trans B 10.1098/rstb.2019.0035; Agudelo-Cantero & Navas 2020 J Therm Biol 10.1016/j.jtherbio.2019.03.010; Rubalcaba & Olalla-Tarraga 2020 10.1111/1365-2656.13181

Results throughout: please supply degrees of freedom with t values.

L285-287: these results do not seem to align with the boxplots provided in Figure 4/3 (page 44). This is probably because of the boxplots showing median not mean, so if possible, please provide a point for the mean of the category overlayed with the boxplot.

L324: an additional limitation of the study is unfortunately the sample size

L342-343: Very high temperatures such as those nearing CTmax would only be beneficial if food, and most importantly water, are not limiting factors – the dependence on water (in context with up to 24% mass loss during <3h of respirometry at high temperatures) needs to be discussed since such high temperatures are also frequently associated with dry conditions. The paragraph of L372-384 touches on water/dry conditions a little, but more could be said.

L398: ‘of’ should be ‘on’

L400-401: sorry, I found this sentence confusing

L416: did you observe discontinuous gas exchange patterns during respirometry or did the ramping temperature prevent discernible patterns from being clearly detected?

Figure 1, 2, and 3 (but labelled figure 4) and Table 1: the order that the life stages appear in as categories along the x-axis of figures and rows in the table appearing in a non-logical life stage order (adult, larvae, pupae or adult, pupae) – please change to larvae, pupae, adult to reflect the logical order of life stages.

Figure 3 (page 46) is not even mentioned in the manuscript. Please make sure it’s referred to and discussed (or leave out or include in a supplement)

Figure 3 or 4 (appears on page 44): the post-hoc letters are either misplaced or the median is very different to the mean report in text (L285-287). From left to right they are a,a,b,b but really look like they should be a,b,a,b from the boxplots because adult F and pupae F are very similar but adult F and adult M are not (yet these adult M/F are apparently not different in terms of means in text). Please check and correct if need be.

Table 1: clarify whether the 95% CL is SD, SE or 95% confidence intervals – all three are mentioned across the methods, table caption and table itself, so it is not obvious what values are actually shown.


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: ESTIMATING THE DIFFERENCES IN CRITICAL THERMAL MAXIMUM AND METABOLIC RATE OF HELICOVERPA PUNCTIGERA (WALLENGREN) (LEPIDOPTERA: NOCTUIDAE) ACROSS LIFE STAGES
Review round: 1
Reviewer: 3

Output:
Basic reporting: I think the framework of the paper could be tightened significantly. The introduction is not particularly well organized, and it makes it hard for the reader (or reviewer) to really understand WHY the study is important, what the main gaps in knowledge are, and especially the theoretical basis of some of the predictions and expectations. I also think they authors should pay special attention to grammatical mistakes (there are many and I have pointed some of them out) and sentence structure. I would like the authors to do a much more thorough search in the literature for work done by numerous scientists addressing many of these issues, instead of citing the same papers over and over again.
Experimental design: No comment.
Validity of the findings: No comment.
Additional comments: The authors report the results of their experiments investigating how thermal physiology (specifically metabolic rate and critical thermal maxima) vary across ontogeny in a moth. I think the topic of the study is important; indeed we know very little about why insect populations are plummeting and certain thermal physiological traits are largely unknown for most insect adults, let alone their juvenile stages. The paper seems well-suited to Peer J’s scope and broad audience. Having said this, I do have a number of issues with the manuscript that I detail below. I mean for these comments to be helpful as the authors revise their manuscript.

MAJOR COMMENTS:

OVERALL: I think the framework of the paper could be tightened significantly. The introduction is not particularly well organized, and it makes it hard for the reader (or reviewer) to really understand WHY the study is important, what the main gaps in knowledge are, and especially the theoretical basis of some of the predictions and expectations. I also think they authors should pay special attention to grammatical mistakes (there are many and I have pointed some of them out) and sentence structure. I would like the authors to do a much more thorough search in the literature for work done by numerous scientists addressing many of these issues, instead of citing the same papers over and over again. Here are just a few papers to pay attention to:

Addo-Bediako, Kimberley Sheldon, Arthur Woods, Sylvain Pincebourde, Raymond Huey, Alisha Shah, Michael Dillon, Sarah Diamond

Addo-Bediako, A., Chown, S. L., & Gaston, K. J. (2000). Thermal tolerance, climatic variability and latitude. Proceedings of the Royal Society of London. Series B: Biological Sciences, 267(1445), 739-745.

Deutsch, C. A., Tewksbury, J. J., Huey, R. B., Sheldon, K. S., Ghalambor, C. K., Haak, D. C., & Martin, P. R. (2008). Impacts of climate warming on terrestrial ectotherms across latitude. Proceedings of the National Academy of Sciences, 105(18), 6668-6672.

Sheldon, K. S., & Tewksbury, J. J. (2014). The impact of seasonality in temperature on thermal tolerance and elevational range size. Ecology, 95(8), 2134-2143.

Diamond, S. E., Nichols, L. M., McCoy, N., Hirsch, C., Pelini, S. L., Sanders, N. J., ... & Dunn, R. R. (2012). A physiological trait‐based approach to predicting the responses of species to experimental climate warming. Ecology, 93(11), 2305-2312.

Shah, A. A., Woods, H. A., Havird, J. C., Encalada, A. C., Flecker, A. S., Funk, W. C., ... & Ghalambor, C. K. (2021). Temperature dependence of metabolic rate in tropical and temperate aquatic insects: Support for the Climate Variability Hypothesis in mayflies but not stoneflies. Global change biology, 27(2), 297-311.

Pincebourde, S., Dillon, M. E., & Woods, H. A. Body size determines the thermal coupling between insects and plant surfaces. Functional Ecology.

These are only a handful of papers that I think have excellent theoretical background and could help the authors revise the framework more succinctly. There are of course many, many more. I continue to detail my comments below by section.

INTRODUCTION:

1) The introduction isn’t as well organized as it could be. In some cases, the beginning of a paragraph does not set up the information in the paragraph. A general rule of thumb is that the first sentence of the paragraph should allude to the theme of that paragraph- also try to stick with one idea per paragraph. For example, in the paragraph starting on Ln 53: The first sentence seems to suggest you will talk about different life stages and why they might differ in CTmax. However, the rest of the paragraph is about how CTmax values are sensitive to methodology.  In addition, please pay attention to the transitions between paragraphs. Each paragraph should have a transitional sentence that prepares the reader for new information from the preceding paragraph. As it stands, many of the paragraphs read like separate, disparate thoughts, one after the other. An example of this is on the paragraph ending at Ln 67 and the one beginning on Ln 68. Perhaps mention plasticity in the previous paragraph so the following one doesn’t seem to be a completely new topic.

2) I feel that introduction is lacking a clear and robust discussion on why different life stages should differ in thermal tolerance (and corresponding citations).

3) Lns 37-38: What exactly is meant by “population dynamics” in this context?  Population dynamics typically refers to the changes in population size as a function of recruitment of young, death,  and migration. I am struggling to see the connection between studying the thermal limits, metabolic rates and population dynamics in the paper, because you are not measuring those dynamics. I would like to see a clear discussion of these ideas to set up the introduction more throughly. The importance of physiology on population dynamics is never mentioned in the Discussion either.

4) Lns 101- 102: I read this sentence as if CTmax is the only determinant of survival for a small insect at extreme temperatures. I disagree. What about biochemical shifts that are temperature-dependant and happen long before CTmax is achieved (see Hotchachka and Somero’s work). Similarly, what about the danger of desiccation, which is linked with temperature tolerance? I caution the authors about writing sentences in such absolutes. All thermal ecologists and physiologists agree that these traits are complex and we still don’t know exactly how they work or affect organisms.

5) Lns 103 - 137: there is certainly a lot of theory about how metabolic rate should change as a function of temperature. There are also a variety of metabolic rate measurements, e.g. resting or basal metabolic rate (which, I think is what you measured). Active MR measures other aspects of an animal’s energy budget. At a minimum, I would like to see a more clear explanation of what type of MR was measured (resting/active), why it was measured, and why you might expect differences to exist between the life stages. I understand that some of the information is already in the manuscript, but a more concise paragraph with these details may help the flow of the introduction.

METHODS:

Ln 172; What temperature were the eggs kept at?

Ln 177: I did not know you could clean lepidopteran eggs this way!

Ln 218: Should this whole sentence be bolded instead of just parts of it?

I think you used a clever way of standardizing CTmax measurements across life stages. Nice.

Note that word “data” is plural so please write, “data are”, “data were”, not “data is” or data “was”.

Ln 246: do you mean “life stages”?

If you are not dealing with mass with CTmax, you could provide the F statistic and p value for CTmax regressed over mass to show it was non-significant.
I am surprised that you get such strong results with such a small sample size. Am I correct that you only measured 5 larvae?

Lns 16-17: I would say “upper thermal limit”. Tolerance is typically a range, whereas a limit is a set point(s), which is what you are referring to in your paper.

Ln 37: delete “moving”. Do you mean shifting the dynamics? or simply the dynamics (which implies variation and change).

Ln 38: “impact” is used twice in the same sentence. Revise.

Ln 45-46: Limits -> limit. Missing “ are” before commonly.

Ln 99: Should say “pupates”

Ln 102: Comma after e.g.

Ln 128-128: Should say “performs”

Lns 140-141: Decide between “thermophysiology” and “thermal physiology”. I personally prefer the latter and believe it is grammatically more accurate. Regardless, pick one and be consistent.

Ln 255: I think you mean Tukey.

RESULTS:

The figure numbers do not match the figure references in the results. Please go through it carefully. For example, you never mention Figure 3 in the main text.

Figures 3 & 4 caption numbers are switched.

I would like to see sample sizes somewhere in the graphs, captions, or in a separate table.

Figure 3: Are pupal CTmax values actually different from the adult values? If they were not part of the analysis, then why are they shown together on the same plot?

DISCUSSION:

Lns: 296-297: You could cite many other much more recent studies on global insect declines.

Ln 317: change to “evolutionarily”

Ln 321-324: I am not sure I understand your explanation. Are you saying that because there is low genetic diversity among lab-reared and wild populations, you expect thermal physiology to be similar between the two groups? I don’t think this is necessarily true. Genetics is not all that accounts for differences in physiology, as you have mentioned elsewhere in your paper. Think about thermal history, plasticity, trade-offs, etc.

Ln 335-337: To me it seems logical that you would expect thermal physiological differences between adult and larval stages. On one hand, larvae are so much more sedentary and likely have a much harder time thermoregulating via behavior, so one could argue they should have a higher tolerance to heat stress. Adults, on the other hand, are far more mobile and could be better-suited to seeking preferred temperatures. This might result in the evolution of lower heat tolerance in adults compared to larvae.

Lns 346-348: Can you say this a little more clearly? I don’t understand how “microclimatic conditions… are higher”. Do you mean that the maximum temperatures are higher? If so, how is that possible? If you have data to back up your statement, I think it would be good to put it here.

Lns 354-355: Indeed, your data suggest that males and females do not differ in their upper thermal limits during short term exposure. But what about long-term exposure or long-term effects on fecundity?


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: ESTIMATING THE DIFFERENCES IN CRITICAL THERMAL MAXIMUM AND METABOLIC RATE OF HELICOVERPA PUNCTIGERA (WALLENGREN) (LEPIDOPTERA: NOCTUIDAE) ACROSS LIFE STAGES
Review round: 2
Reviewer: 1

Output:
Basic reporting: no further comment on original
Experimental design: no further comment on original
Validity of the findings: no further comment on original
Additional comments: The authors have extensively revised their manuscript in response to the numerous comments from all reviewers. The revised version has been improved greatly. Responses to some questions raised by reviewers were not elaborated on much, but they have at least made a substantial effort to revise the manuscript text, figures, and supporting information in line with the reviewer comments.

I have some new minor comments:

L89-92: this sentence presents three or four very disjointed ideas in it and it itself seems out of place. The content is fine, but it needs to be separated into smaller sentences to make sure the links among them are logical.

Figure 3: the outlying Q10 35-45˚C value for pupae that is apparently above 10 is surely unrealistic when the remaining pupae estimates are around 1-2… please check the data and even if it appears technically sound, consider carefully whether such a value is biologically realistic. I really doubt it is.

Also, a few small editorial suggestions:

L39: change ‘an’ to ‘that an’

L70: change ‘ground surface as well as on plant surface’ to ‘ground and plant surfaces’

L71: change ‘the many microclimatic conditions including’ to ‘substantial microclimate variability, including’

L370: change ‘have’ to ‘has’


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: NO EVIDENCE OF EFFECTS OR INTERACTION BETWEEN THE WIDELY USED HERBICIDE, GLYPHOSATE, AND A COMMON PARASITE IN BUMBLE BEES
Review round: 1
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The effect of glyphosate on non-target organisms, especially on pollinating insects, is an international research hotspot. Although many papers have been published on its effect on honeybees, there are few studies on its effect on bumblebees.This paper studies the acute and chronic toxicity of glyphosate to bumblebees and its interaction with parasites, filling the gaps in the research on the effects of glyphosate on bumblebees. The paper still has the following  parts that need to be revised:
1. Line 122  one of "2010"should be deleted.
2.Line 180 "."should be added after"excluded".
3.Line 478 it is better to add some expression after"sublethal effects", for example add "(Sucrose/Glyphosate consumption,Parasite intensity".
4.Are the worker bees used to conduct the experiment randomly selected? Is there a uniform age or not, and why?


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: NO EVIDENCE OF EFFECTS OR INTERACTION BETWEEN THE WIDELY USED HERBICIDE, GLYPHOSATE, AND A COMMON PARASITE IN BUMBLE BEES
Review round: 1
Reviewer: 2

Output:
Basic reporting: The article meets the basic criteria regarding writing and citations and is well structured. Figures are easy to interpret.
Experimental design: The experimental design is appropiate, following OECD protocols and modifying them when needed.

I would have added a PER assay testing for learning could provide a more detailed insight about what could be happening in a field scenario, where bumblebees need to forage.

I have a question regarding parasite examination of the mother colonies: Why you haven't used a molecular determination approach instead the faeces parasite count? I think this way you will be missing some parasites. If you are only looking for Nosema, Crihitida and trypanosomatids, you will miss viruses. Please clarify this in the materials and methods section.
Validity of the findings: As the authors claim, more replicates will be needed in future studies in order to test Gly effects. But I think this is valid approximation.

Discussion section
Line 515-518: Although co formulants do have an effect per se, in fields where GLY is appled, bees are in contact with the formulation, and not with the active ingredient alone. If your intention is to work with a realistic scenario, I think your research could be benefited in future studies from using field formulations.
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: NO EVIDENCE OF EFFECTS OR INTERACTION BETWEEN THE WIDELY USED HERBICIDE, GLYPHOSATE, AND A COMMON PARASITE IN BUMBLE BEES
Review round: 1
Reviewer: 3

Output:
Basic reporting: The work conducted by Straw & Brawn seems to be a very carefully conducted series of in vivo experiments following regulatory guidelines to investigate the effects of a commonly used herbicide, glyphosate, and a parasite, C. bombi, on bumble bees. They found no clear evidence for detrimental effects of glyphosate, C. bombi, or the combination of both on bumble bees. Experimental conditions and statistical analyses are described in detail, although the writing is relatively dense sometimes.

I could see that the authors thought carefully about their experimental conditions and the results obtained, as most of the questions I had were promptly answered in the discussion, such as the differences in parasite load between experiments which could have led to different outcomes across experiments.

Below I provide very minor comments as an attempt to improve the quality of the study.

1.	The words “parasite” and “pathogen” are used interchangeably throughout the manuscript, but they are not synonymous. Please revise this.
2.	The figure numbers do not follow the order they appear in the main text.
3.	Line 310 – I think you cited the wrong figure. It should be figure 8, instead of 9.
4.	Line 690 – please double check the percentages. It is weird that a 50% reduction in reproductive success is not significant.
5.	Figure 1 – Include in the legend whether these concentrations come from nectar or pollen sources.
6.	Also, there are several citations for unpublished work or work under review. Hopefully, they will be published by the time this study is accepted for publication, and if so, don’t forget to make the proper modifications.
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: LANDSCAPE AND MICROHABITAT FEATURES DETERMINE SMALL MAMMAL ABUNDANCE IN FOREST PATCHES IN AGRICULTURAL LANDSCAPES
Review round: 1
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: The meaning and significance of this study should be further stressed and clarified.
Validity of the findings: no comment
Additional comments: 1. Why do you choose to study these two species in this area? Needs more clarification.
2. The meaning and significance of this study should be further stressed and clarified in the Introduction and Discussion part.
3. Conclusions should be more concise, and detailed information may be moved to results and discussion part.
4. In the abstract, context and background should be stated before stating "We studied the populations of two small mammals (Apodemus agrarius, A. sylvaticus) in 19 wood remnants along a gradient of agricultural landscape intensification in Friuli Venezia Giulia (north-eastern 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: LANDSCAPE AND MICROHABITAT FEATURES DETERMINE SMALL MAMMAL ABUNDANCE IN FOREST PATCHES IN AGRICULTURAL LANDSCAPES
Review round: 1
Reviewer: 2

Output:
Basic reporting: See comments below. Generally very good.
Experimental design: Very good.
Validity of the findings: Very good. See minor comments below.
Additional comments: General comments
Globally there are concerns about anthropogenic land-use change effects on biodiversity. In this study, the authors investigated how landscape and microhabitat features determine small mammal abundance (two Apodemus spp.) in forest patches in agricultural landscapes. The work makes an important contribution to our understanding of small mammal species persistence in anthropogenic landscapes.

I suggest replacing ‘due to’ with ‘because of’ throughout.

Specific comments
Title: Fine.

Abstract:
L24 Insert a general sentence.
L25 Maybe say how many woodlots and over what time.

Introduction:
A good synthesis of the relevant literature.
Avoid one-sentence paragraphs, so combine 2nd para with 3rd.
L65 ‘responded’.
L72 Correct author’s name ‘Bricker’.
L82 Replace ‘proved to’ with ‘shown’.
L97 Both references are not in the list.
L108 What were your predictions?

Methods:
Good explanation of methods used.
L116 Insert Latin names.
L214 Put GIS in full at 1st mention.
L214 Is it not QGis?
L218 Correct ‘polygon’.
L234 Change to ‘analyses’.

Results:
Were both species caught in all your sampling sessions?
L291, 294 Avoid one-sentence paragraphs so combine.
L299, 318 Insert space ’Table 2’.
L301 Check Journal format for the presentation of statistics.
L315 ‘3.2’.

Discussion:
Perhaps a bit long.
L518 ‘Conclusions’.
L523, 557 ‘showed’.

References:
I have not checked if the Journal format was followed. Most in-text references are listed.
Please check and follow the Journal format. Some Journal titles are abbreviated, and others not. Some Journal titles do not have each word starting with a capital.


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: LANDSCAPE AND MICROHABITAT FEATURES DETERMINE SMALL MAMMAL ABUNDANCE IN FOREST PATCHES IN AGRICULTURAL LANDSCAPES
Review round: 1
Reviewer: 3

Output:
Basic reporting: The article is very comprehensive in terms of content and contains all relevant sections. However, I have the following concerns:
1. Some sections (e.g., Materials and methods) contain many grammatical and typographical errors. I have highlighted some of them for the authors' consideration and revision.
2. There are many long sentences (e.g. Lines 551-556) which makes it hard to read.
3. The introduction should be improved to ensure a proper logical flow. For example, there is a sudden change in grammatical tense on Lines 65 and 66. There are also redundant phrases that need to be removed. For example, "on their mosaics" on Line 106.
4. The references are very relevant, but I did not find any recent ones. I noticed that the latest reference is nearly 4 (i.e. 2017) years old. The authors will agree with me that there have been several advances (i.e., important studies) during this period that need to be referenced to improve the quality of the introduction, discussion, and conclusions.
5. I also noted with concern the variable weird names such as "N_spec_arbr", "Dmt_med_fst". The authors should endeavor improve this.
6. There are other editorial revisions that will improve the quality of the paper. For example, a better rendering of the equation on Line 191 with an equation number. I have highlighted most of these editorial errors for authors' review and revision.
7. Table and figure labels are too long in my view. Authors may want to shorten them.
Experimental design: The methodology is sound. no comment
Validity of the findings: No comment
Additional comments: Please see the attached annotated PDF for details.


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: LANDSCAPE AND MICROHABITAT FEATURES DETERMINE SMALL MAMMAL ABUNDANCE IN FOREST PATCHES IN AGRICULTURAL LANDSCAPES
Review round: 2
Reviewer: 1

Output:
Basic reporting: Clear and good.
Experimental design: Clear and good.
Validity of the findings: Good.
Additional comments: Revisions have been made according to previous review report, and I think it could be accepted.


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: LANDSCAPE AND MICROHABITAT FEATURES DETERMINE SMALL MAMMAL ABUNDANCE IN FOREST PATCHES IN AGRICULTURAL LANDSCAPES
Review round: 2
Reviewer: 2

Output:
Basic reporting: See below
Experimental design: See below
Validity of the findings: See below
Additional comments: I thank the authors for their detailed revision. They have addressed all my concerns and I find the manuscript acceptable now.


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: LANDSCAPE AND MICROHABITAT FEATURES DETERMINE SMALL MAMMAL ABUNDANCE IN FOREST PATCHES IN AGRICULTURAL LANDSCAPES
Review round: 2
Reviewer: 3

Output:
Basic reporting: Minor editorial modifications aimed at improving the readability of the paper have been made for authors' consideration.
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The modified manuscript is attached for authors' consideration


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

Input:
Title: LANDSCAPE AND MICROHABITAT FEATURES DETERMINE SMALL MAMMAL ABUNDANCE IN FOREST PATCHES IN AGRICULTURAL LANDSCAPES
Review round: 3
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: I thank the authors for their hard work on this paper. Congratulations and best wishes.


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: PERFORMANCE IN MORTALITY PREDICTION OF SAPS 3 AND MPM-III SCORES AMONG ADULT PATIENTS ADMITTED TO THE ICU OF A PRIVATE TERTIARY REFERRAL HOSPITAL IN TANZANIA: A RETROSPECTIVE COHORT STUDY
Review round: 1
Reviewer: 1

Output:
Basic reporting: Kassam et al. report on a small single-centre experience with approximately 300 intensive care patients. The aim of the paper is to compare SAPS 3 and MPM 0-III for outcome prediction.
Experimental design: Retrospective cohort study.
Validity of the findings: From my point of view, there is no novelty in this study.

(e.g. https://pubmed.ncbi.nlm.nih.gov/28523584/)
Additional comments: This retrospective analysis does not generate new hypotheses.


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: PERFORMANCE IN MORTALITY PREDICTION OF SAPS 3 AND MPM-III SCORES AMONG ADULT PATIENTS ADMITTED TO THE ICU OF A PRIVATE TERTIARY REFERRAL HOSPITAL IN TANZANIA: A RETROSPECTIVE COHORT STUDY
Review round: 1
Reviewer: 2

Output:
Basic reporting: The article is mostly written clearly and unambiguously; the phrase "mortality predictive performance" - placed prominently in the title - does seem a bit odd, though. I would suggest phrasing such as "performance in mortality prediction for ease and clarity.

Literature references provided are sufficient to provide adequate background and context.

The manuscript follows the usualarticle structure; figures, tables are well presented; raw data are shared.

The article overall is self-contained with relevant results to hypotheses.

Specific comments:
Line 160ff: Please provide absolute numbers in addition to percentages for clean reporting.

Line 112ff: The usual acronym is "SAPS 3" (rather than "SAPS III") and should be used consistently.
Experimental design: The article includes original primary research with aims and scope I would consider within the stated width of the journal. The research question well defined, relevant and meaningful. It is clearly stated how this research fills an identified knowledge gap.

The investigation has been performed to a technical standard that requires clarification. Ethical standards are adhered to. Methods used are described with insufficient detail as of yet.

Specific comments:
Line 122: "The Intensive care unit (ICU) of the Aga Khan Hospital is 15 bed modern and well-equipped unit which provides level III services to all kind of critically ill patients."- While this is undoubtedly the case, "modern" and "well-equipped" are both ill-defined terms which should not be used in such an absolute manner.

Line 127: "The ICU is run by a multidisciplinary team, comprising of the primary care physician [...]" - Does that indeed mean a primary care physician (i.e., doctors specialising in primary care) or physicians of the primary specialty of care of the respective patient (e.g., surgeons), as in an "open" ICU model? Please clarify.

Line 133ff: "A 133 total of 747 adults patients were admitted to the ICU from August 2018 to April 2020. A sample size of 331 patients was determined to be sufficient to give the study a 80% power and 95% confidence for  detection of 10% difference in performance between SAPS-III and MPM0-III." Such a power-calculation is laudable, but larger sample size - especially if available - would certainly be preferable. Indeed, the sample size may be sufficient for score comparison, but is undoubtedly weak for performance assessment of scores in general (without the risk of over- or underfitting).

Line 148ff: I miss any definition of mortality (i.e., the study's endpoint) in this section. Later paragraphs only adress "survivors" and "non-survivors". Line 59 of the hints at "in- intensive care mortality". If that is the case, this either constitutes a flaw in the study's design or needs special attention, as all models cited actually calculare hospital mortality rates rather than ICU mortality rates (with the latter necessarily being lower).

Line 155ff: "A Hosmer-Lemeshow goodness-of-fit test was used to assess the calibration of the models with a p-value > 0.05 considered statistically significant." This is confusing and contrary to my understanding of the Hosmer-Lemeshow-test. Furthermore, the known limitations of this very test must be expressed and emphasised.
Validity of the findings: Line 95: "APACHE-IV has long been considered more precise for predicting mortality than the other scoring systems" - This statement cannot be supported by the given reference 13 (Kuzniewicz et al.), since it does not adress SAPS 3 (but rather SAPS II, which - no matter the name - has been derived independently from SAPS II).

Line 101ff:  "External validation in other ICU populations reported that SAPS 3 had good discrimination, but poorer calibration, when compared with APACHE-IV and MPM0-III. However, it may have greater potential for international use since the score was derived from data in more than one country" - While the latter is certainly true, the statement overall cannot hold true, if calibration were off overall. Recalibration for regional or local patients cohorts would be required.

Line 200ff: "Calibration of each scoring system exhibited good performance. The goodness of fit Hosmer-Lemeshow test and p value of each scoring system is shown in Table 4 below." I suspect the listed Chi-Square statistics were calculated according to the 'original' proposed calculation of Ĉ (i.e., with nine degrees of freedom). This should, however, be clearly stated.

Line 215ff: I suspect the calculation of mortality with SAPS 3 was performed using the formulae presented in the original publication. This needs to be emphasised, as the authors themselves point out, that multiple recalibrations exist. More importantly, it is hospital mortality that is calculated - its comparison with ICU mortality is in itself a fallacy.
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: PERFORMANCE IN MORTALITY PREDICTION OF SAPS 3 AND MPM-III SCORES AMONG ADULT PATIENTS ADMITTED TO THE ICU OF A PRIVATE TERTIARY REFERRAL HOSPITAL IN TANZANIA: A RETROSPECTIVE COHORT STUDY
Review round: 2
Reviewer: 1

Output:
Basic reporting: Issues of reporting remain, e.g. "Most of the patients were admitted to the ICU from the emergency department 306(92.5%)" which should probably read "Most of the patients were admitted to the ICU from the emergency department (n=306, 92.5%)" or similar. Changes like these can be performed by the authors at the editors discretion without further need for peer review. Similarly, some final English editing is advised.
Experimental design: The methodology now seems sufficiently described.
Validity of the findings: "Our results highlight the impact of prolonged length of Stay (LoS) in the ICU which is associated with higher adjusted odds of mortality among critically ill patients." I would advise to refrain from such statements without the utilisation of time-to-event utilisation. Moreover, this statement and the analyses leading up to it do not seem to adress the stated research question.
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: NIGHTTIME MIGRATIONS AND BEHAVIORAL PATTERNS OF PEMPHERIS SCHWENKII
Review round: 1
Reviewer: 1

Output:
Basic reporting: Re: Manuscript 62502

General comments:
The paper aligns with the intent of PeerJ

The subject of the paper is of interest and the type of data presented on nocturnal fish are rare.

The combination of field and tank experiments was sensible.  However, substantial revision is required.

In general, there was poor utilisation of the data available. It was clear that data were collected and mentioned somewhat casually in the text of the results, but we’re not presented in figures or tables.  I have made a number of suggestions.

Some editorial work required, especially with respect to word usage.

In general, the captions to the figures were incomplete.

Specific comments
Use past tense for material and methods and results.

Materials and methods
Visibility of tag (by distance?) and duration the tag held its glow? State how the tag was attached to the peduncle?
Line 105 and 138  – re mapping – what about landmarks such as large rocks and like?
Line 146 re: 200 fish, 60 Pempheris – what were the rest?

Results
Line 157 before starting the field.
Line 171 – 172 – fish spent about 2 minute in in-reef then to out reef, but n lines 178-180 – they usually spent time in shallow water – clarify!
Line 172 re: (Mean 2.0) give SE and sample size
Line 174, give percentage of time as an individual and as a group?
Line 176, Seriously without SCUBA how mapping to 30m and even more unlikely tracking to 150 m achieved?
Line 178 - they usually spent time in shallow water? This is unclear from the maps. If lots of individuals tracks provide time versus depth budgets in a table with all replicates
Lines 181- 189 - variation in trackers with time not presented as table # of graphs.

Figures
Fig 1 – provide an estimate of size of fish and the size of the tag. The tags are attached with a cable tie?
Fig 2 needs a scale
Fig 3 caption - specify for individuals A-D.  There is a need for a table that includes all individuals – e.g., time spent at inner and outer areas of reefs and rate of movement/hr or minute.
No need to present depths as minus values
Fig 4 sexes of fish in the spawning image?
Figure 5 Feeding behaviour may be best presented as a line - from essentially sunset to sunrise? In the caption Specify the number of nights that this plot was developed from and the number of fish.
So site 3 and 2 were not used!?

Discussion
Line 216 “chase” = follow?
Lines 219-223 – these are results!
Line 229 - 238 inference that may spawn at outer reefs - delete as no evidence for this.
Lines 238 to 248, comment - spawning may not occur because stressed in aquarium, or not the right environmental triggers. Sampling frequency did not allow for the possibility that spawning only happened on so nights (e.g., semilunar).
Line 249 Annese & Kingsford found quite different behaviour by species. One species was found in the shallows in groups, while the other was rather solitary and stayed in deeper waters
Line 255 - Re migration several kilometres - little evidence of this.  An hourly rate of movement was calculated, but chances are they spent most of the night foraging once the reached a targeted area?
Lines 307 - 326, any conclusions are made that do not relate to the results. In fact, little is said here on the nature of migrations - which is what the paper is about!
Examples of errors Wrong use of words:
Line 25 – ‘nocturnal fish group’ should be ‘nocturnal group of fish’
Line 35 – re: “on surface”  you mean near the surface?  Correct throughout.
Line 109 and elsewhere.  “In-reef” and “out-reef” would be better as inner reef and outer reef, change throughout
Line 126 ‘despising’; line 128 were not was; line 147 ‘comparted’;
Line 161 re: gleamed” you mean there was still ambient light?
Line 164, waiting for and individual to lead the migration; line 301 lowers caps for polychaeta.
In-reef and out-reef = inner and outer reefs.
Experimental design: all comments are in the first box
Validity of the findings: all comments are in the first box
Additional comments: all comments are in the first box


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: NIGHTTIME MIGRATIONS AND BEHAVIORAL PATTERNS OF PEMPHERIS SCHWENKII
Review round: 1
Reviewer: 2

Output:
Basic reporting: Many lapses in English and poor choice of wording.  It really needs another thorough revision to improve the English and choice of words.  "Chasing" is used inappropriately.  This should be changed to "Following" as we hope that the diver observing fish movements is not actually chasing the fish, which surely would affect its movements.  In therms of the behavioural category "swaying"... this is not a term that is currently in use.  Is there an accepted behavioural term that could be substituted here.

The literature is largely out of date and not used to generate any hypotheses.
Experimental design: Research questions were not clearly defined.The basic sampling design is really far to minimal to make quantitative assessment possible.  This is really just a collection of anecdotal observations.
Validity of the findings: The use of the luminescent tags is probably novel.  But any likely effects of the tags on behaviour were not assessed.  The sample size was too low and I do not think it is appraise to publish "rough" back of the envelope estimates of movement distances.  This can only be improved by further work.
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: CONSTRUCT VALIDITY AND INTERNAL CONSISTENCY OF THE BREAST INFLAMMATORY SYMPTOM SEVERITY INDEX IN LACTATING MOTHERS WITH INFLAMMATORY BREAST CONDITIONS
Review round: 1
Reviewer: 1

Output:
Basic reporting: Thank you for the opportunity to read this important paper on the development of an instrument to follow the progress of inflammatory symptoms of the breast during lactation.  The paper is well written and the language is clear and professional. Whilst I acknowledge that this is a methodological paper (and possibly part of a future doctoral thesis) I feel that the information contained in the background is insufficient for this stand- alone paper to be properly understood by the reader. I have included some comments and suggestions regarding this below.
Introduction
The research that is cited in the background is sparse and requires a broader international perspective. The authors are surely aware that there is a history of dissent within the scientific community as to whether conditions such as mastitis are inflammatory or infectious and because this paper clearly states the measurement of inflammation as the driving force for the instrument’s development, it would be advantageous to acknowledge the fact that researchers have viewed this differently through the last 3 decades and it would enhance the understanding of the paper, if the reader was provided with references to the latest scientific approach to understanding the possible pathways of inflammatory symptoms of the breast. Here are some suggestions for literature that might be included to improve the international perspective and to give weight to the paper: (Ingman et al 2014, Martin et al 2007, Kvist et al 2008, Delgado 2008, Collado et al 2009, Hunt et al 2011, Jahanfar et al, 2013, La Tuga et al, 2014 and Human Microbiome FAQ produced by the American Academy of Microbiology).

In an international perspective, it is unusual that breastfeeding women are treated at physiotherapy clinics. The setting of this study and the background to why physiotherapists have the role of lactation consultants in Australia should be clarified, either in the background or the methods section.

On line 69 the reader is told that more than 400 clinicians are already using the instrument but without knowledge of the total number of clinicians, this statistic has no meaning.

The aim of the study is clearly stated and also that there is no existing instrument to measure ICLB. You might point out how useful such an instrument will be for future efforts to carry out international research on ICLB . This will take knowledge about ICLB forward at an increased pace. (You could include this in the conclusion or as a rationale for the research question in the introduction).
Experimental design: Materials and methods
The materials and methods are clearly presented.
On line 128, “unreported responses” seems to be an oxymoron. Does this mean that there was no answer? If so, these are not  “responses”.
Lines 102-103 state that the search for eligible case notes was continued until 160 were identified. How did the authors decide on this number of case notes?
Lines 111-112 states that “the statistical analysis required a minimum of 10 to 20 scores per assessment item”. This is unclear but may just be a question of semantics; a minimum is usually given as one figure and not as a range.
Lines 151 – 153 states that ICLB has been viewed as a continuum and Betzold 2007 is cited. I contend that there is no robust empirical evidence for a continuum for ICLB. The idea of a continuum has been contested (Kvist et al., 2008). WHO (Mastitis: causes and management) suggests that breast abscess seems also to occur spontaneously, irrespective of previous mastitis, and therefore questions the idea of a continuum. This is an important point because practitioners who believe that there is a continuum will be apt to prescribe antibiotics early, to stop progress of the condition (which they would expect if there was a continuum). We are fighting a global battle against antibiotic resistance and we have a responsibility as researchers to not perpetuate unnecessary use of antibiotics.
Validity of the findings: Results
The results of the analyses appear robust and the raw data is unambiguous.
Lines 181 to 187 are headed “Analysis” and give an account of how the material was standardised. This section probably belongs to the Materials and Methods section.
Line 192: I find it odd that the items Fever, Ache and Headache are included under a factor called “Wellness” (although in the table the item is called Wellness/Sickness). These would indicate illness rather than wellness. This should be considered and changed throughout the manuscript. This is particularly visible in Table 3 where the factor is “Wellness” and the item description contains three signs of illness.

Discussion
A very nice and well-balanced discussion.
Additional comments: An important and well designed study although limited by the collection of material from only one clinic.


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: CONSTRUCT VALIDITY AND INTERNAL CONSISTENCY OF THE BREAST INFLAMMATORY SYMPTOM SEVERITY INDEX IN LACTATING MOTHERS WITH INFLAMMATORY BREAST CONDITIONS
Review round: 1
Reviewer: 2

Output:
Basic reporting: No comments
Experimental design: No comments
Validity of the findings: No comments
Additional comments: The authors have done a lot of analytical work to determine the Breast Inflammatory Symptom Severity Index in lactating mothers with inflammatory breast conditions. It is wonderful. But I do not understand the purpose of this study and the practical significance of the results obtained. What are the recommendations for using this index for patients with mastitis? If it is just for making a diagnosis, then the symptoms of mastitis are so specific that it does not require the use of any severity scales. The main problem with mastitis is the rapid progression of the disease into a purulent form. At the same time, the deterioration often does not correlate with the severity of the symptoms. Therefore, the managing of mastitis (prescribing antibiotics, surgical drainage) is often determined not by the severity of symptoms, but by the time from the onset of the disease and the resolve symptoms against the background of treatment. Unfortunately, the authors did not conduct such an analysis (correlation between the index and the stage of mastitis, as well as the effectiveness of therapy).


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: CONSTRUCT VALIDITY AND INTERNAL CONSISTENCY OF THE BREAST INFLAMMATORY SYMPTOM SEVERITY INDEX IN LACTATING MOTHERS WITH INFLAMMATORY BREAST CONDITIONS
Review round: 2
Reviewer: 1

Output:
Basic reporting: No comment.
Experimental design: No comment.
Validity of the findings: No comment.
Additional comments: Congratulations on a very nice paper.


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

Input:
Title: COMPARATIVE ANALYSIS OF TWELVE MITOGENOMES OF CALISCELIDAE (HEMIPTERA: FULGOROMORPHA) AND THEIR PHYLOGENETIC IMPLICATIONS
Review round: 1
Reviewer: 1

Output:
Basic reporting: L. 83: How does mtDNA contribute to the control of pests?
L.104: It would be better if the authors can provide the information for longitude and latitude.
L.149: PhyloSuite has been published in Molecular Ecology Resources, please cite the up-to-date citation, not the Biorxiv preprint.
L. 180: 1000 ultrafast bootstraps is too few. The authors should use at least 10,000, but you may also use 100,000.
L. 216: Please italicize gene names throughout the MS.
Experimental design: L.174: Did the authors directly align the nucleotide sequences of PCGs without considering the codon frame? PCGs should be aligned in codon mode; you may either use PhyloSuite or MEGA for that.
Validity of the findings: L.240-242: Confirm this, normally AT skew of insects is positive.
L.251-L.252: Please also confirm the skew here.
L. 311: 100 is the max value of the bootstrap support. What is the meaning of “BPs>100” here?
L. 315-318: Peltonotellini was not the basal lineage.
L. 318-319: Augilini doesn’t form a sister group with Caliscelini and Peltonotellini.
L. 422-430: The two clades that the authors described as “top” and “bottom” of the tree respectively were sister-clades, so describing them as top and bottom is wrong.
L.435-436: In the phylogenetic analysis, the authors only included members of this family (except for outgroups), so they can’t conclude this.
Additional comments: The authors present 12 new mitogenomes for Caliscelidae species, the data themselves contribute much to the phylogeny or molecular studies for these insect pests. I only have few concerns regarding the M&M and the interpretation of the phylogenetic tree.

I cannot see Figure 2 anywhere.

Figure 4: replace Ommatidiotini with Ommatidiotinae?

Please note that the line numbers mentioned here are referring to the track changes file.


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: COMPARATIVE ANALYSIS OF TWELVE MITOGENOMES OF CALISCELIDAE (HEMIPTERA: FULGOROMORPHA) AND THEIR PHYLOGENETIC IMPLICATIONS
Review round: 1
Reviewer: 2

Output:
Basic reporting: no comment
Experimental design: No comment
Validity of the findings: No coment
Additional comments: 1. When authors start the write the result part, they should clear that how many mitogenome studied in this study. How many species dataset prepared for the phylogenetic study.
Studied mitogenomes belong to which family, subfamily and so…..on
Provide the details of mitogenomes which were retrieved from gene bank.
Why authors have used 5 outgroups. Is there any relevance in this study? Explain it

Authors generated 12 mitogenomes data of family Caliscelidae under two subfamilies   Ommatidiotinae (9) Caliscelinae (3).

Supplementary files names (sheets) are not corresponded with text.

Authors has made a confusion that theyy have used 25 mitogenomes or 21 mitogenomes. Somewhere they have mentioned 25 line or in phylogeny 21

Insects are a good model to study the gene arrangement pattern. Here, authors did not discuss the gene arrangement scenario in this family. All the species have conserved gene order or they have some gene arrangement. One paragraph can be added. Authors should compare the Caliscelidae  gene order to ancestor.

Author have generated the mitogenome data of unidentified species Peltonotellus sp.. What is the relevance?


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: COMPARATIVE ANALYSIS OF TWELVE MITOGENOMES OF CALISCELIDAE (HEMIPTERA: FULGOROMORPHA) AND THEIR PHYLOGENETIC IMPLICATIONS
Review round: 1
Reviewer: 3

Output:
Basic reporting: The language is clear and unambiguous. I have no issues with the reporting.
Experimental design: 2. Experimental Design
Between Specimens’ collection and DNA extraction, there was no specific description of sample preparation.

This according to the authors, was given attention. However, it doesn't appear like anything was done. I believe to encourage reproducibility, the steps and procedure observed to prepare the sample for use in DNA extraction should be described.
Questions like how the muscle tissue of each Caliscelidae species was collected and prepared for extraction should be answered.
Validity of the findings: Findings appear to agree with the research hypothesis, together with the data provided. Issue with discussions and conclusion were added to the attached reviewer's response document
Additional comments: Additional comments have been added to the attached reviewer's response document.


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: COMPARATIVE ANALYSIS OF TWELVE MITOGENOMES OF CALISCELIDAE (HEMIPTERA: FULGOROMORPHA) AND THEIR PHYLOGENETIC IMPLICATIONS
Review round: 2
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: The authors have resolved most of my concerns, however, there still remain some mistakes and some places need further clarification:

1. If the authors didn’t align PCGs in the codon mode, it is a drawback with respect to their phylogenetic analysis. I suggest that authors should re-align PCGs in the codon mode, and construct the phylogenetic tree anew.

2. Regarding the skew, please make sure that you didn’t confuse the two strands.

3. The authors still seem to have misunderstood several parts of the phylogenetic tree. A. You can’t say that Peltonotellini was the basal lineage of the phylogenetic tree, even disregarding the outgroups. B. Augilini formed sister-group with the clade containing Peltonotellini and Caliscelini, so they exhibit equal position. Claiming that Augilini is in the upper part of the tree is wrong. C. Claiming that Yotuus is located in the lower part of the Augilini clade is incorrect. You can say that Yotuus was basal to the Augilini.
Additional comments: no comment


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

Input:
Title: COMPARATIVE ANALYSIS OF TWELVE MITOGENOMES OF CALISCELIDAE (HEMIPTERA: FULGOROMORPHA) AND THEIR PHYLOGENETIC IMPLICATIONS
Review round: 2
Reviewer: 2

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


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

Input:
Title: COMPARATIVE ANALYSIS OF TWELVE MITOGENOMES OF CALISCELIDAE (HEMIPTERA: FULGOROMORPHA) AND THEIR PHYLOGENETIC IMPLICATIONS
Review round: 3
Reviewer: 1

Output:
Basic reporting: The authors present 12 new Caliscelidae mitogenomes and conduct phylogenetic analysis using 16 Caliscelidae species. They resolved most of my concerns, I commend the authors’ effort to obtain and sequence as many species as they could (12) as opposed to sequencing one mitogenome at a time. The results and discussions are generally sound now, and I would like to recommend this manuscript for publication in PeerJ after resolving the following two minor problems:
1. line 364: Peltonotellus (Peltonotellini) is not located at the middle of the tree, imaging that if you rotate the node containing Calisselinae and Ommatidiotinae, Peltonotellus can locate at the top of the tree.
2. line 377: delete ";" after the word mitogenomes.
Experimental design: no comment
Validity of the findings: no comment
Additional comments: no comment


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

Input:
Title: IDENTIFICATION OF THE GOLDEN-2-LIKE TRANSCRIPTION FACTORS GENE FAMILY IN GOSSYPIUM HIRSUTUM
Review round: 1
Reviewer: 1

Output:
Basic reporting: Zhao et al described the identification of GLK genes in Gossypium hirsutum using structural genomics approaches and discussed their potential functions in drought stress based on tissue-specific expression patterns, drought response (qPCR), and co-expression networks. The reviewer found that the methods and results are solid and may require only minor revisions.
Experimental design: The authors analyzed the GLK gene family in cotton using bioinformatics approaches. These analyses can help to understand gene function. This is good work, and well written.
Validity of the findings: For my part, this paper is prepared well. The authors used well-established methods to perform analysis. Results and data interpretation are comprehensive and reasonable. The conclusion is also a suggestive conclusion. However, there also some concerns that should be addressed before publication.
Additional comments: A professional English editor is needed to improve the language. Grammar mistakes and misspellings are mostly in the manuscript, which makes the manuscript difficult to read. The authors should check the manuscript very carefully before the submission; well I understand that English is not their native language.


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: IDENTIFICATION OF THE GOLDEN-2-LIKE TRANSCRIPTION FACTORS GENE FAMILY IN GOSSYPIUM HIRSUTUM
Review round: 1
Reviewer: 2

Output:
Basic reporting: The general outline of the manuscript obeys the journal conditions, so to that far, the line is OK, however, the flow of thought is not proper, and it is paramount for the authors to polish the manuscript before resubmitting for a second evaluation.
Experimental design: Materials and methods
Databases: to be corrected to the database, only a single database is shown in the manuscript. Moreover, this section should be merged with “the genome-wide identification of the GLK genes in Gossypium hirsutum” .
The section for genomewide identification…. Should be corrected to state “Identification of the GLK genes in Gossypium hirsutum”
A serious concern of the use of the two protein domain “PF00249.31 and PF14379.6” in which PF00249.31 Myb_DNA-binding (PF00249) - Pfam: Family and PF14379.6: Myb_CC_LHEQLE; MYB-CC type transfactor, LHEQLE motif, and yet the gene or the TF family under study is the GLK: can the authors explain the mismatch, because this raises serious question of the identification of the gene or the TF under study.
Phylogenetic tree analysis:  The authors have used protein sequences obtained from A. thaliana, tomato and G. hirsutum. The very first question which rings into the mind of the reader, what was the justification of using the protein sequences from these three plants to carry out the phylogenetic tree analysis, moreover, is tomato evolutionary closer to cotton compared to Theobroma cacao. The second serious concern is no information is provided within the manuscripts on how the protein sequences from Tomato and Arabidopsis were obtained.
This section is poorly done and must be redone, with protein sequences obtained from amore closer relatives to cotton.
Promoter cis-element analysis of GLK:  Change to Cis-regulatory elements. Is true as pointed by the authors that through the determination of the cis-regulatory elements one is able to determine the function of the gene or the TF, the answer is no, so the authors need to rephrase.
Differential gene expression analysis: The authors need to simply state “RNA seq expression analysis” reason for doing this not stated not indicated, and furthermore, it is secondary data already in the public domain, unless the data was generated by the authors, this need to be clearly shown in order to improve the validity and novelty of the submitted manuscript
Stress treatments and qRT-PCR analysis:  The treatment initiated “cold, salt and drought” and yet the RNA seq used as the reference point were profiled after the plants were exposed to “low temperature, salt and drought”, the question to the authors can low temperature be equated to cold stress?
Validity of the findings: Not valid, because in the materials and methods, serious questions have been raised, the results will only be valid if all the issues raised corrected.
Additional comments: Substantial revision is needed before the paper could be reviewed again, and if the authors are not able to carry out all the issues stated, I fully recommend rejection. Moreover, the manuscript is too shallow, if possible the authors could be advised to validate the key genes or the TFs as per the RNA seq. and RT-qPCR analysis.


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: POTENTIAL LINEAGE TRANSMISSION WITHIN THE ACTIVE MICROBIOTA OF THE EGGS AND THE NAUPLII OF THE SHRIMP LITOPENAEUS STYLIROSTRIS: POSSIBLE INFLUENCE OF THE REARING WATER AND MORE
Review round: 1
Reviewer: 1

Output:
Basic reporting: The study by Giraud and colleagues focuses on the active fraction of eggs and nauplii microbiomes of Litopenaeus stylitostris, also with the aim of understanding the role of the environment in conferring microorganisms, as well as the potential vertical and horizontal transmission of microbial taxa. The manuscript is overall novel, well written, in professional and unambiguous language, and the scientific aims and conclusions are well reported and discussed; the topic of this study is certainly of interest for the readership of this Journal. I only suggest some minor comments, described in more detail below.
Experimental design: I suggest the Authors to add some clarifications on the study design; for instance, it would be helpful to have a scheme (also as Supplementary Material) of the tanks system (ResI, ResII, filtration sizes etc). At lines 137-139 I suggest to add references for the packages used to run analyses.
Line 159: It would be useful to have an idea of the range of ASVs found in the analyzed samples, for example by adding min-max ASVs values at the end of this sentence.
Line 175: Plancomycetes
Lines 234-248: In my opinion, this paragraph it’s excessively long and could be moved in the M&M/results sections.
Validity of the findings: Results and conclusions are well supported and discussed. I only suggest to remove L403-405 from the Conclusions paragraph.
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: POTENTIAL LINEAGE TRANSMISSION WITHIN THE ACTIVE MICROBIOTA OF THE EGGS AND THE NAUPLII OF THE SHRIMP LITOPENAEUS STYLIROSTRIS: POSSIBLE INFLUENCE OF THE REARING WATER AND MORE
Review round: 1
Reviewer: 2

Output:
Basic reporting: 1. The structure of the manuscript follows the journal standards.

2. There are some grammatical, linguistic and spelling mistakes that should be revised.

3. The authors are recommended to re-check Peer J authors instructions about the reference guidelines

4. I could not access the raw data, please check the accession number. No results were found in NCBI SRA repository for the following submissions “Submission ID SUB9828953, BioProject ID PRJNA736535”
Experimental design: There are few major issues with the experimental design that prevents from completely supporting the claims and conclusions made in the manuscript.

Please provide more information’s/details about the samplings (time points, sample size, numbers of replicates, water sample volumes filtrated) in all experiments (M1, M3, M4). In the current version is not clear which type of samples have been collected in each experiment. Please also describe the way the nauplii have been collected.

Furthermore, please justify the use of 2 extraction kits.

Please explain why the Eggs and nauplii sequenced twice (Lines 164 – 165)

I advice the authors to give some information’s to the readers about the ASVs clustering at the materials and methods section.
Validity of the findings: 1. The normalisation method it is not needed to be mentioned in the conclusion section.

2. There are also recent studies suggesting vertical transmission in farmed fish eggs and are worth mentioning

3. The results are relevant results to hypotheses.
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: POTENTIAL LINEAGE TRANSMISSION WITHIN THE ACTIVE MICROBIOTA OF THE EGGS AND THE NAUPLII OF THE SHRIMP LITOPENAEUS STYLIROSTRIS: POSSIBLE INFLUENCE OF THE REARING WATER AND MORE
Review round: 1
Reviewer: 3

Output:
Basic reporting: The manuscript is globally well written although I have a few comments:
- from line 344 to 353: this section lacks structure and some sentences are hard to understand (e.g. line 344). I would suggest to rewrite this section and avoid starting sentences with "As for...". Also, in line 352, a reference is needed for the characterization of Rothia in vent shrimp eggs.
-line 364: spelling mistake "beneficial bacteria" not "benefic bacteria"
Also it's either "potential beneficial bacteria" or "a potential beneficial bacterium".
-line 141. The authors mentioned a submission ID and BioProject ID. However, no raw data is available under these codes. Please make sure the raw data have been released in the NCBI SRA repository.
Experimental design: Replicates have only been used for the last experiment (M4). In addition, there are no samples from the water in tanks when collecting eggs and nauplii samples. It is likely that the bacterial composition of the seawater in tanks is different from the reservoirs, in particular following the addition of EDTA. As such, there could be bacteria in the seawater of tanks that were not detected in the seawater of the reservoirs, potentially overestimating the number of specific ASVs found uniquely in the eggs and nauplii. This is an important bias for the generation of the Venn diagrams which most results in this study are based on.  While this issue is mentioned in l302, it could have been avoided by collecting seawater samples during the collection of eggs and nauplii.
Validity of the findings: As it is currently described in the legends and in the main manuscript, it sounds like the bar graphs (in Figures 3, 4, and 5) represent the relative abundance of specific ASVs in relation to all ASVs. However, they actually represent the relative abundance of a subset of ASVs (those selected from the Venn diagram). The actual relative abundance of these ASVs is far less than what is displayed in the bar graphs since many ASVs are excluded for the generation of these graphs. This needs to be better explained in the manuscript and the figure legends.
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: PRELIMINARY PLACEMENT AND NEW RECORDS OF AN OVERLOOKED AMAZONIAN TREE, CHRISTIANA MENNEGAE (MALVACEAE)
Review round: 1
Reviewer: 1

Output:
Basic reporting: see below
Experimental design: see below
Validity of the findings: see below
Additional comments: I believe the manuscript would benefit from a narrower focus. The manuscript seems to be making several different arguments and I would be tempted to jettison the first of the two main arguments 1) The authors argue that in the various recent compilations of data on Brazilian plants, some taxa have been overlooked. (This is, in some respects, trivial). 2) Among the overlooked taxa is Christiana mennegae and not only do the authors add it to the Brazilian flora (again), they also sequence it and place it in a phylogenetic context, and they look at the trichome morphology of the same species. In addition, they extend its range to Amazonian Peru.

The argument about collecting gaps and taxa being overlooked is based on one example. If the authors are to do justice to this argument, they need to marshal additional data otherwise their argument comes across as anecdotal.

Title. The “identity” of Christiana mennegae is not something that is or was in question and the word “identity” should be suppressed. Usually when a taxonomic paper mentions “identity”, the plant in question is or was grossly misinterpreted (e.g., placed in the wrong family).

The “new records” for Brazil, if they consist only of Amazonas and Pará collections, are not new. These state records were reported already. What is new, is the extension of the range of the species to Amazonian Peru.

Pantropical – I think of Pantropical taxa as occurring in the Americas, Africa, and Asia. A species that is found in the American and African tropics only is not pantropical.

Distinguishing the American species of Chirstiana – The text is confused. This information could be expressed well as either a revised key (following Kubitzki, 1995) or a table.

Description of trichomes. I am not sure that multi-angulate captures what you are trying to convey. When I look at your images, I see some stellate hairs with the arms (comparatively few in number) in a single plane (rotate) and others with many more arms in several planes. I would describe these latter as tufted, rather than angulate.

References. These need to be checked carefully. There are many spelling errors and incomplete citations. Also, the punctuation is not standardized. Some titles are consistently lowercase while others are mixed case, etc.

Illustrations. The illustrations are all well done and informative.


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: PRELIMINARY PLACEMENT AND NEW RECORDS OF AN OVERLOOKED AMAZONIAN TREE, CHRISTIANA MENNEGAE (MALVACEAE)
Review round: 1
Reviewer: 2

Output:
Basic reporting: No comment.
Experimental design: No comment.
Validity of the findings: No comment.
Additional comments: The authors of this study explore the taxonomy, micromorphology, and phylogenetic relationships of poorly known species, Christiana mennegae, in an almost equally poorly known genus of Malvaceae. This is a short paper.  It includes new observations and so certainly makes a contribution to our knowledge of this species.  However, the paper seems a little insubstantial and important information is missing from the methods and results.  The editor needs to make the decision whether this study is significant enough to warrant publication.  The paper is very clearly written, although there are several minor problems with the English (grammar and idiomatic expressions).  I strongly encourage the authors to find a good English-speaking editor to help.  More detailed comments are below.

Intro:

The authors begin by making a compelling argument that knowledge of the flora of the Amazon region is incomplete and that additional studies, and new kinds of studies, are needed.  This is nice writing, but this subject is not revisited in the rest of the paper.  How does more information on one species help our understanding of the Amazon region?  How does this study illustrate and fix the general problem?  What strategies would prevent other species from being similarly overlooked in the Flora of Brazil?  Do they predict other species will have similar enormous range expansions if/when there is more coordination about taxonomists?

The paper would benefit from a clear summary of the genus Christiana in the introduction.  How many species are in the genus, their distribution, generic limits, any question or investigation into its monophyly.  All of the information is scattered in the manuscript, but it needs to be presented clearly in the introduction.  Also, if Christiana africana has the same distribution as Ceiba pentandra (tropical America and tropical Africa), it should not be called pantropical.  The authors may want to track down Kubitzki’s treatment of Malvaceae in the 2003 Families and Genera of Vascular Plants.

For the taxonomic treatment, how many specimens were seen (either physical specimens or digital images)?  How many new collections were made?  Fig. 1 shows four dots.  What does each dot represent?  A collection?  A population?  Was the map prepared only using herbarium specimens, or are new collections included?  Can the authors make estimates of population sizes based on their field work?  If the species is dioceious, how many staminate specimens were observed, and how many carpellate?

How was conservation status determined?

SEM: There is no information on how SEM was performed.  Were structures reconstituted from dry specimens?  And if so, how?  How were the mounts prepared?  Did they go through a dehydration series?  Were they sputter-coated?  Was the SEM under full vacuum or low vacuum?  What structures were examined with SEM, and why?  In the methods, the authors state “vegetative and reproductive structures,” which includes pretty much the entire plant.  But in the results, they only discuss trichomes and stomata.   Was there an a priori reason to examine trichomes and stomata, or were these the only observable traits?  The micrographs in Fig. 3 are beautiful, but there is so little known of the micromorphology of Brownlowioideae that is is difficult to interpret the significance of these results.  How many specimens
 were examined, and how many structures (leaves, flowers, fruit) per specimen?  Were leaves mature or immature?  All of this information is important to evaluate how robust the observations are.

Phylogenetic analysis: The authors add two short sequences (rbcL and ITS2) from C. mennegae to a previously published dataset of Malvaceae.  There are several issues here.  It would be good to see a hypothesis or rationale for performing a phylogenetic analysis.  Is there a reason to think that Christiana is not monophyletic?  That C. mennegae is not a member of Brownlowioideae?  The dataset that the authors added their sequences to is enormous, and far larger than they need (unless there’s a possibility that C. mennegae is not in Brownlowioideae).  Hernandez-Gutierrez and Magallon reanalyzed data that had been previously published by other authors, so it is not accurate to say that they published sequences from 239 species of Malvaceae.  I don’t know the details of this matrix, but I’m sure from the description that there is a large amount of missing data.  None of that is reported here.  My suggestions: State your hypothesis for the phylogenetic analysis. Construct a matrix that is a reasonable test of your hypothesis.  This probably means Brownloidoideae and a few outgroups from other subfamilies of Malvaceae.  Exclude gene regions for which there are large amounts of missing data.  Give more detail on the final dataset, your analyses, and the results (seven lines for the results is a red flag).  I appreciate the authors’ caution in interpreting their phylogenetic results.  But, given that C. mennegae and C. africana appear to only have the slow-evolving rbcL sequences in common, it is quite remarkable that their trees show these two species as sister taxa.  Their monophyly is not strongly supported, but I would not expect it to be given the data.

Figures:

The Fig. 1 legend needs more detail.  How many collections is this map based on?  Do these represent individual collections, known populations?  We need some numbers here.  The compass and scale bars would be more readable if they were also in white.

Fig. 2 is lovely, but the legend should include information of the specimen that the illustration is based on.

Fig. 3 is also lovely, but also needs specimen information. I don’t like the colorization, and I think it makes it difficult to see detail, especially of the stomata and glandular trichomes.  It helps a little, perhaps, in Fig. 3a, but not at all in b-h.

Fig 4: See my comments on the phylogenetic analysis.

Fig. 5 Again, lovely, but are the colors enhanced in c-e?  If so, it should be stated.  Information on this particular specimen should be given.  I don’t think you need both Fig. 5d and 5e, and the cropping of 5e is problematic.


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: PRELIMINARY PLACEMENT AND NEW RECORDS OF AN OVERLOOKED AMAZONIAN TREE, CHRISTIANA MENNEGAE (MALVACEAE)
Review round: 2
Reviewer: 1

Output:
Basic reporting: See below
Experimental design: See below
Validity of the findings: See below
Additional comments: I read the revised manuscript and the authors’ response to points the two reviewers raised in their reviews. While the authors addressed the easily resolvable problems, they avoided the more serious ones. The current title promises “phylogenetic placement” of Christiana mennegae while the abstract and introduction suggest that the study addresses large scale biodiversity and digitization issues. The manuscript does not deliver on these promises. A single molecular sequence for an infrequently-collected species as well as some original morphological (SEM) observations are presented. While there is value in these observations, they are “preliminary.”


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: PRELIMINARY PLACEMENT AND NEW RECORDS OF AN OVERLOOKED AMAZONIAN TREE, CHRISTIANA MENNEGAE (MALVACEAE)
Review round: 3
Reviewer: 1

Output:
Basic reporting: I only have suggestions in order to improve the content and correct minor errors.

Some suggestions are made below:
-	Line 37: Include “Brownloioideae” in keywords.
-	Line 62: The citation for De Candolle (1824) must be included in the References.
-	Line 74: “Secco (2000) recorded Asterophorum mennegae…”. Include after “Asterophorum mennegae” the term “(= C. mennegae)”?
-	Line 112-113: “It was not necessary to critical-point dry them.” I think that this sentence is not necessary because the authors previously mentioned on line 109 that the samples were dried.
-	Line 186-187: The mentioned Table includes results from this manuscript, but also results compiled from the literature. The results obtained from the literature must be indicated and referenced.
-	Line 207: This information must be transferred to M&M, not mentioned here.
-	Line 227-231: The description of trichomes is in Supplementary Material, but it’s important to note the types of trichomes in this paragraph.
-	Line 228: It’s not necessary repeat the citation of “Theobald et al. 1979.” This information is already noted in M&M.
-	Line 245, end of Results: Figure 5 wasn't explored. In the absence of information about this figure, I suggest transferring it to Supplementary Material.
-	Line 263: The two cited authors must be included in the REFERENCES.
-	Line 276-277: Glandular trichomes were also seen in the petioles. See Figure 3A. The description of glandular trichomes must also be included in the Supplementary Material.
-	Legend, Figure 2: In C, it's difficult to see the mentioned trichomes. I suggest an insert like Figure F.
-	Legend, Figure 3: Figure B mentioned the presence of stomata, but this structure wasn’t mentioned in the Results.
-	Legend, Figure 3: In E, the magnification of the image makes it impossible to see the type of trichome.
-	Legend, Figure 3: In F, it would be appropriate to provide an image with the same magnification as that in Figure 3D. In the presented figure, it is impossible to view the trichome type.
-	Legend, Figure 3: Arrows cannot be seen in the figure.
-	In the Electronic Supplementary Material:
o	Material and Methods: In the first paragraph, line 4, include in the References the citation of Ramalho et al. 2018.
o	Results: In Table 1, include the source of the information not original to this manuscript. The data about the geographic distribution of all cited species must also be included.
o	Micromorphology study: first paragraph –
	Line 1-3: The glandular trichomes weren’t mentioned or even described.
	Line 4-5: The stellate-rotate trichomes aren’t visible in Figure 3B. “fig 3C” should be deleted.
	Line 8-9: Figures 3A-C are about vegetative structures (leaf and petiole), not reproductive. The reproductive structure is represented in Figures 3D-F.
	Second paragraph: How is this text related to “Micromorphology study”?
Experimental design: no comment
Validity of the findings: no comment
Additional comments: I would like to congratulate the authors for their research. The core of the article is creative and obtained from data that could not even be considered.

The images presented are of high quality, and the analyses were performed equally well.