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: EVIDENCE FOR A NON-LINEAR CARBON ACCUMULATION PATTERN ALONG AN ALPINE GLACIER RETREAT CHRONOSEQUENCE IN NORTHERN ITALY
Review round: 1
Reviewer: 2

Output:
Basic reporting: The manuscript is clearly written in good English, with one or two places noted for improvement in the author comments below. Overall the authors include sufficient background and I point out a few places where clarification or more context could be used. The article structure is clear and logical, and the hypotheses for the study are well-defined and clearly related to the results.
In the discussion and introduction it might be interesting to cite some of the ideas regarding soil C development and storage potential brought by (Janzen 2006, Bradford et al. 2016, He et al. 2016).

References:
Bradford, M. A., W. R. Wieder, G. B. Bonan, N. Fierer, P. A. Raymond, and T. W. Crowther. 2016. Managing uncertainty in soil carbon feedbacks to climate change. Nature Climate Change 6:751–758.
He, Y., S. E. Trumbore, M. S. Torn, J. W. Harden, L. J. S. Vaughn, S. D. Allison, and J. T. Randerson. 2016. Radiocarbon constraints imply reduced carbon uptake by soils during the 21st century. Science 353:1419:1424.
Janzen, H. H. 2006. The soil carbon dilemma: Shall we hoard it or use it? Soil Biology and Biochemistry 38:419–424.
Experimental design: The experimental design is adequate with clearly defined and interesting research questions. The investigation is performed to a high standard and the methods and results are clearly presented. In the author comments I point out some ideas for minor improvements.
Validity of the findings: The data are well presented and the findings are placed in context with a relevant discussion of the meaning and implications. I give a few suggestions and more detailed comments in the author comments, below.

I am curious whether there are specific projections for glacial retreat in the Matschertal? If yes, this could be mentioned to provide context for the expected rate of change.

The data are provided with the exception of September fluxes, which seem to be missing from the excel document. In figure S1 I think it would be helpful to show the current glacier extent in relation to the transects. For the data reported in tabular form explicit metadata would be helpful. This can be short and just a description of columns and abbreviations.
Additional comments: Line 53-54: could be more clearly/directly phrased, eg: Glacial expansion in the Alps peaked around 1860 and since then have been retreating leaving an abundance of bare ground…

Line 72-74: does this study measure changes in energy balance? This sentence could be deleted. It is an interesting fact that is not totally relevant to the C cycle focus of the study.

Line 75: use a direct object. What does ‘this phenomenon’ refer to? The phenomenon of glacial retreat or of the radiative forcing discussed in the previous sentence?

Line 81: why the mention of black carbon here?

Line 92-97: this is nice.

Line 102: I think this would read more smoothly if changed to: ‘the variation in carbon and nitrogen is considered to influence photosynthesis and respiration equally’.

Line 103-111: I’m not sure I totally understand the mechanisms for prevailing soil processes? Is that because microbial communities and soil C&N cycling might have a stronger influence on C dynamics than the vegetation? This paragraph talks a lot about plants, it might be better to focus on the soil?

Line 112-118: scenario 3 is laid out very nicely, focused on reasons why plant processes could be the most dominant mechanism for C accumulation.

Line 139: That’s a nice map. Would it be possible to indicate the current glacial extent as well? Maybe a dotted line? Or perhaps a google earth image with T1-T3 indicated for reference?

Line 212: the data provided in the supplement is missing September? I found only July and August. The data is well organized and generally easy to understand. Still, I would having a metadata page that describes the columns and some values. For instance, what are 2b,3b,etc and ‘ck’? One of those is presumably a repeat with no filter. I understand that f0-f11 are increasing filter thickness (decreasing light).
Why was the zero-filter measurement repeated again at the end? I think typically we assume that full photosynthesis might take some time to recover which is why we measure from full to low-light conditions.

Line 192-194: oh, that’s a cool method!

Line 214: modified how? I am not sure what modification would be needed to the light-response method?

Line 227: September data are missing in supplement?

Line 244: This is a nested design, with 5 chambers nested within each transect. Was a nested analysis (mixed effects model) tested? The sample size is quite small so it might be difficult but a mixed model might have greater statistical power because it could account for presumably greater similarity between collars from a single transect.

Line 266-268: are these three numbers presumably per transect? Reported as: T1, T2, T3? Please clarify.

Line 269: earlier stage of succession is T1? Add to text.

Line 280: alfa – should this be the Greek letter alpha? What does alpha signify?

Line 282-283: what are conoid and LIA moraine? Transects?

Line 285: this is a nice way to report transect T1, T2, T3 numbers.

Line 286: was the vegetation already senesced in September?

Line 299-303: conoid and LIA moraine again?

Lime 350-354: were the regressions done on fluxes normalized by light and temperature?

Line 357: could the terms LIA and conoid be explained? Many readers might not know these terms since this is not a journal for glaciologists. And could the terms be linked explicitly to T1, T2, T3?

Line 390: The comment on forest establishment needs a little more context. Is temperature the primary limitation? Do dispersal and soil development play a role in limiting trees?

Line 397: presumably the relationship between BG biomass and soil present a feed-back. As more soil develops more roots can grow, which leads to more soil formation?

Line 423-428: this sentence is a little hard to follow. It might be better to break it up into two, and fix some grammatical errors.


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: LEVELS OF TOOTHACHE-RELATED INTERESTS OF GOOGLE AND YOUTUBE USERS FROM DEVELOPED AND DEVELOPING COUNTRIES OVER TIME
Review round: 1
Reviewer: 1

Output:
Basic reporting: The article falls below standard English usage in both style and grammar. At various places, it is difficult to make sense of the manuscript. Also, the manuscript does not flow well and it is not an easy read.

The manuscript is sufficiently supported by literature, however the details presented in the manuscript does not support the overall context well.

Figures and other data are presented well. Raw data excel file is missing some information.

Results are relevant to hypothesis but the hypothesis itself is not very informative.
Experimental design: Methods are fairly well laid out but the utility of multiple testing to largely look at trends of usage of a keyword does not provide an interesting view of the problem and therefore conclusions are restrictive.
Validity of the findings: There seems to be little impact of the results in the overall field of data science relevant to user interests of data platforms for questions relevant to dentistry.

The discussion of the manuscript is informative and brings out relevant points but the remaining article does not dissect the topic in detail. While I understand that this is restricted by data available from Google trends, the information reviewed and presented contributes little to the advancement of the topic.
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: LEVELS OF TOOTHACHE-RELATED INTERESTS OF GOOGLE AND YOUTUBE USERS FROM DEVELOPED AND DEVELOPING COUNTRIES OVER TIME
Review round: 1
Reviewer: 2

Output:
Basic reporting: Interesting study and relevant to current scenario
Experimental design: Abstract:
Lines # 40-42: Background: Authors have mentioned “In this sense, hypothetically, source-based patterns of health related Web searches might differ between individuals from developed and developing countries, due to their educational backgrounds.” Differences could be due to broader social, economic, political, cultural, psycho-social reasons, not just educational backgrounds. The authors can consider rephrasing this sentence.

Line # 46: Methods: “This longitudinal study analyzed……” Methodology section of main text indicates that this is a “retrospective longitudinal study”. The latter is more appropriate.

Main text:
Line # 64: Introduction: “The spreading of Communication and Information Technologies (ICTs) stimulated individuals in…..”. Consider changing “Communication and Information Technologies” to “Information and Communication Technologies”, as indicated in the parenthesis by the authors
Line # 104: Search term included was “Toothache-Disease”. Individuals might use many other search terms/key words while using the internet to know about tooth ache.
Lines # 110-112: “Developed countries were composed by Australia, Chile, Japan, United Kingdom, and United States, while developing countries were composed by Brazil, Mexico, Russia, Saudi Arabia and South Africa”. Was this classification based on World Bank, United Nations or any such agencies? Kindly include reference for the same.
Lines # 110-112: Among the countries included, notable exceptions are China and India. Can authors provide any explanation for the same?
Lines # 110-112: Can the authors provide actual information about “availability and penetration of the Internet” in these countries? It will enhance the value of the findings of the present study.
Lines # 112-115: “To avoid bias related to the selection and comparability of countries, both groups were statistically similar in relation to burden of untreated dental caries in permanent teeth, characterized by toothache (Institute for Health Metrics and Evaluation, 2018).” Can authors present the results in tabular form and explain what “statistically similar” is?
Lines # 110-112: The usual reference cited for global burden of disease is World Health Organization (WHO) databank. Can the authors explain why or how Institute for Health Metrics and Evaluation, 2018 provides greater/better data than WHO?
Lines # 110-112: Untreated dental caries need not always present itself as toothache. Can the authors make a note of this in the discussion section?
Validity of the findings: Results:
Table 2 can be presented as first table. Can the authors provide descriptive explanation of their findings before presenting RSV trends? They can inform the reader about mean Google and YouTube searches for developed and developing nations.

Can the authors consider exploring intragroup comparisons of outcomes? It would be interesting to observe for a statistically significant increase in the number of searches in Google and YouTube within the 2 groups.

Discussion:
Internet usage may not necessarily translate into initiation of remedial or corrective measures. Authors can consider adding the same in discussion section
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: LEVELS OF TOOTHACHE-RELATED INTERESTS OF GOOGLE AND YOUTUBE USERS FROM DEVELOPED AND DEVELOPING COUNTRIES OVER TIME
Review round: 1
Reviewer: 3

Output:
Basic reporting: None.
Experimental design: None
Validity of the findings: None
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: LEVELS OF TOOTHACHE-RELATED INTERESTS OF GOOGLE AND YOUTUBE USERS FROM DEVELOPED AND DEVELOPING COUNTRIES OVER TIME
Review round: 2
Reviewer: 1

Output:
Basic reporting: Nocomment
Experimental design: Corrections done
Validity of the findings: Relevant
Additional comments: Can 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: DIVERGENT WATER SOURCES OF THREE DOMINANT PLANT SPECIES FOLLOWING PRECIPITATION EVENTS IN ENCLOSED AND MOWING GRASSLAND STEPPES
Review round: 1
Reviewer: 1

Output:
Basic reporting: General Comments
The study entitled, ‘Divergent water sources of three dominant plants at precipitation events in natural and mowing grassland of a typical steppe’ attempted to determine water sources of three dominant plant species growing in mowed and natural steppes. The method of cryogenic vacuum extraction of water, which was used in the study, is one of the most widely used and accurate extraction methods to obtain plant and soil water samples for δ18O. This task is daunting, but produces valuable data. Thus, the method used is very appropriate.

The study has great potential, especially in informing restoration ecologists on possible choices of plant species for different niches. However, the handling of the data and its analysis could have been slightly different. I have now highlighted these suggestions in the results section of this report and also as comments on the attached PDF version of the manuscript.

Language: The manuscript has several grammatical mistakes that need attention in order to improve on clarity. I have highlighted some of these in the manuscript.
Title: Firstly, I suggest revision of the title to ‘Divergent water sources of three dominant plant species following precipitation events in natural and mowed grassland steppes’.
Abstract: see corrected pdf text for many specific comments
Introduction: I suggest removal of the first sentence (line 36-37) as it is not adding value to the manuscript.
Line 62-63 Provide a more accurate description of the study sites. This is not an accurate description of the ecosystems used in the study.  'coastal' not 'riverbanks' Mediterranean-type deserts' not ‘temperate steppes'
Line 64 Its not true, many studies done in South Africa are in semi-arid areas
Line 81 The sentence is too long and wordy. Consider rephrasing it
Methods:
Line 101 What is the relevance of accumulated temperature? It gives an impression of heat storage.
Line 106 You have only introduced, 1. Natural grassland and 2. Mowing grassland. ‘Experimental grassland’ will confuse readers. Rather refer to experimental area
Line 114 Rephrase for clarity. Do you mean that: Two precipitation events were recorded in July 2016, which are 10.8mm on the 29th of July and 20.0mm on the 30th of July.
Line 114 Rephrase to ‘Experimental data were analyzed using SPSS Version 19.0 (SPSS Inc., Chicago, USA)’
Line 153 rephrase to ‘which are 1) surface water layer (0-5cm), 2) shallow water layer (5-10), 3) middle layer (10-40) and 4) deep water layer.’
Line 154 Change ‘The’ to ‘An’
Line 155 Provide the source of the model. Include in parenthesis the source.
Results:
Revise figures as follows:
Figs 1 & 2 Because four depth categories were mentioned in line 153 (0-5; 5-10; 10-40, >40) , this figure should be redrawn to show average moisture values of these categories, possibly on a secondary axis or as a different fig with the categories plotted against δ18O value and moisture contents. Then relate species δ18O values to soil layer (0-5; 5-10; 10-40, >40) averages. Then conclusions around soil layer would make more meaning to the readers
Figure 3 Refer to ‘three plant species’ not ‘3 plants’. It implies that you used three individual plants, one of each species.
Figure 4 Symbols are missing. This is making your results difficult to understand. eg. What do different lines stand for? What does a circle or a triangle mean?
Line 162 This is difficult to understand. The authors mention a ‘very significant change’ but ‘no significant effect was detected’? What 'effect' was measured here?
Line 165 This is poorly constructed. You mention that 'changes were different' but we know that a change is a difference between two states/levels etc. Please rephrase
Line 167-169 This section needs careful description. I suggest making reference to the four categories that were identified earlier in line 153. What was the purpose of separating soil zones if interpretation is made based on all individual distances measured.
Line 183-184 I think one cannot use ‘largest δ18O value’ if there were no significant differences observed in δ18O values. Please rephrase to correctly to relay the intended message.
Line 186 Delete the word ‘separately’
Line 202-208 Indicate the proportions used in this section.
Discussion
I suggest that you consider revising the current discussion of your results after considering δ18O values for each layer 0-5; 5-10; 10-40 and >40. These δ18O values for soil layers are the potential water sources that should be present in the non-photosynthetic parts of the plant. I think this is still possible with the Isosource model. Signatures of precipitation events (2 rain events) may not adequately describe soil zones explored for water uptake.
Line 214 I suggest that you rephrase this. It suggests lack of confidence in your statistical results. Should be ' were no significant differences between mowed and enclosed grasslands’.
Line 216 Pay attention to your grammar. Use past tense in this section. Rephrase. I could not follow the statement here. Same applies to line 222-225
Line 266 How is shown from your data?
Experimental design: see previous comments in pdf version
Validity of the findings: A revision is suggested after considering suggested comments
Additional comments: see previous 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: DIVERGENT WATER SOURCES OF THREE DOMINANT PLANT SPECIES FOLLOWING PRECIPITATION EVENTS IN ENCLOSED AND MOWING GRASSLAND STEPPES
Review round: 1
Reviewer: 2

Output:
Basic reporting: Line 19, delete extra space;

Line 28, "distribution pattern" is vague, precipitation distribution?;

Line 37, this paragraph of "Utilization of different water sources by plants.." is not very relevant to this study, as this study is not actually characterizing the water sources but only the depth of soil layer for water uptake.

Line 51, this example, "In the Sequoia sempervirens forest.." is very extreme because those trees are very tall and at high elevation. And is not appropriate as an example for this study because plants involving in this study are perennial grasses and shrubs.

Line 61-64, again, authors should focus on expanding the Introduction discussing the gaps in those few studies with steppe ecosystems. Mentioning temporal forests is not useful for this study because they are different ecosystems.

Change captions for table 1 and 2 to "ANOVA tables for ....."
Experimental design: Line 99, add "China"

Line 107, "The experiment was started..";

Line 112, "of each year"? how long was mowing period or only in 2011

Line 122, ''..and combined as one replicate"
Validity of the findings: Line 162-163, I am confused by this sentence. soil water content had a significant change but then no significant effects were detected? so the change before and after precipitation is not significant?

Line 167, should be more specific, at what depth the soil water content started to decrease. Same here in line 169, at what depth there is a reverse?

Line 178, it might be better to calculate % change instead of saying a rapid decrease and a mild decrease.

Line 187-193, I think the results for those isotope should not just be values because they are hard to understand. Authors should give some topic sentences say what those ranges mean comparing among three species.

Line 214, if the effect is not significant, either there is no sufficient data or more variables (factors) should be considered during the experiment. Authors should not explain the results assuming it is significant. Again, in line 229-230, if the result is not significant, why still have implications? what kind of implications? or authors should pick the experimental site with longer mowing history.

Line 235, were there any recent references to be compared with? this paper (and is the only reference here) was from 1983.

Line 240, "In our study, in shallower soil, ...."

Line 244, Since authors discussed the time intervals between the two collections (first date vs. fifth date) below, the example of reference should also be the intervals between sample collections instead of the time intervals between precipitation events.

Line 260, omit "(Yang et al., 2011)"

Need a conclusion section? if this is required by the journal.
Additional comments: This paper reported changes in soil moisture at different depths in mowing vs. enclosed grassland before and after precipitation events. It also studied the oxygen stable isotope ratios of soil water and stem water of three plant species: Leymus chinensis, Stipa grandis and Cleistogenes squarrosa. This paper over discussed some results for which the statistical analyses were actually not significant. Introduction and discussion need more work, please see the above 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: DIVERGENT WATER SOURCES OF THREE DOMINANT PLANT SPECIES FOLLOWING PRECIPITATION EVENTS IN ENCLOSED AND MOWING GRASSLAND STEPPES
Review round: 1
Reviewer: 3

Output:
Basic reporting: This manuscript aims to explore how the three coexisting dominant species Stipa grandis, Leymus chinensis and Cleistogenes squarrosa in a typical steppe community respond to summer precipitation event in water use under two grassland utilization modes (mowing and enclosure). I think that the manuscript addresses interesting questions. However, the manuscript was not well written. There are too many grammatical errors, and the manuscript should be improved greatly in English expression.

I have some important concerns and suggestions about this study.

The abstract should be rewritten because of some grammatical errors.
Line 24 “S. grandis from ……..” should be “S. grandis took up water from ……..”
Line 27 “a more sensitive soil moisture change” should be “a more sensitive change of soil moisture”
Line 29 “the observed differences” should be “the differences”
Line 30-31 “Plant root systems……..”. This sentence doesn't make sense.

You should give a better background of oxygen stable isotope ratios (δ18O) of soil water in introduction section. Why you choose oxygen stable isotope ratios (δ18O) of soil water to explain divergent water sources of three dominant plants, and why you choose Stipa grandis, Leymus chinensis and C. squarrosa as research objects, please increase the explanation and provide references.
Line 60 “the studies on” should be “studies about”
Line 61 “the ecosystems of” can be deleted.
Line 63 “the coasts” should be “coasts”
Line 68 “the semi-arid steppe ecosystem” should be “semi-arid steppe ecosystems”.
Line 69 “the soil water” should be “soil water”
I think there are too many “the” in this manuscript.
Line 79 “changes in” should be “changes of”
Line 81-84 “Therefore……..” should be “Therefore, understanding the mechanisms of plant water use is essential to steppe management by exploring how dominant plants respond to instantaneous precipitation events and use available water resources in natural and mowing conditions .
Line 87 “event” should be “events”
Line 2,18,28,88 The expression about utilization modes should be consistent. “mowing and natural” in the title, “native (in enclosure) and mowing” and “mowing and enclosed grassland” in the abstract, “mowing and enclosure” in introduction section.
Line 88 “did this by detecting” should be “determined”
Line 90-91 “mowing zone and enclosure zone of the steppe” should be “mowing and enclosure plots”
Line 92-94 English language

The methods section needs a lot of improvements, because there are many aspects that are missing or unclear.
Line 100 “a mean annual temperature of -0.4 °C”. Please check the data “-0.4 °C” and provide the year range of calculating the mean annual temperature. I think a mean annual temperature in this site should be about 2°C .
Line 101 “the average plant growth period” should be “the average period of plant growth”
Line 102 “around 200 mm ~ 350 mm” should be “between 200 to 350mm”
Line 105 “or Calcic-orthic Aridisol” should be “equivalent to Calcic-orthic Aridsoils”
Line 106 “The experimental grassland was” should be “The vegetations in the study region are ”
Line 107-109 “The experiment……..” should be “”The enclosure and mowing plots were set up in 2011 to study the effects of annual mowing on native steppe.”
Line 109-111 “A total……..” should be “eight 20 m×30 experimental plots (four enclosure and four mowing plots) were established with a distance 2m between any two plots.”
Line 114-116 “Two precipitation……..” should be “a light precipitation event of 10.8 mm on 29 July and a medium precipitation event of 20.0 mm on 30 July.
Line 118-119 “Plant samples ……..” should be “Non-photosynthetic tissues of plant from the interface”

In results section, you should reanalyze your data.
Line 161 “Effects of ……..” should be “Effects of summer precipitation on soil water content in mowing and enclosure steppe”
Line 163 “but no significant effect was detected (Table 1, Fig. 1)”, I think there were significant effects of summer precipitation on soil water content of 5-10 cm and 10-20cm in mowing and 5-10 cm in enclosure steppe. Soil water content after precipitation was higher than before precipitation.
Line 170-171 “Effects of ……..” should be “Effects of summer precipitation onδ18O of soil water in mowing and enclosure steppe”. You should focus on the differences between two plots. There were significant differences inδ18O of soil water 0-5 cm and 5-10cm in enclosure steppe before and after precipitation events.

There are some inappropriate expressions in Note of Table 3. It is easy to make the reader confused. You should explain “~” in Note of Table 3.
There was no legend in Fig. 4.

References
The references should be in the same format.

Overall, I think this MS should be revised to reach the standard of publication in this journal.
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: DIVERGENT WATER SOURCES OF THREE DOMINANT PLANT SPECIES FOLLOWING PRECIPITATION EVENTS IN ENCLOSED AND MOWING GRASSLAND STEPPES
Review round: 2
Reviewer: 1

Output:
Basic reporting: None
Experimental design: Line 101, why there is a > 0 degree C next to the accumulated temperature?
Validity of the findings: Line 159, subheading can just be "Results", remove "and analysis"

Line 185, i again vote to have the actual value even if it is larger than 0.05. In ecology, sometime if p values fall into 0.05 and 0.1, we might also see that as a weak significant differences. Because sometimes researchers not seeing a significant difference does not mean there is not, it might be the small sample size or larger variation.

Line 220, change to "were similar between..."

Line 262, not just rainfall intensity, but the land topography (e.g. slope) also impact the soil water distribution.

Line 281-285, this is the conclusion part, so I suggested authors to add the details of how they define "middle, deep soil layer" by giving the range of depths.
Additional comments: I appreciated authors' efforts to revise this paper. I think this paper can be accepted after some minor edits suggested above.


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

Input:
Title: FLY PALAEO-EVO-DEVO: IMMATURE STAGES OF BIBIONOMORPHAN DIPTERANS IN BALTIC AND BITTERFELD AMBER
Review round: 1
Reviewer: 1

Output:
Basic reporting: The paper is an interesting, relevant and novel contribution pointing out an important shortcoming in the research in amber fossils: the lack of attention to fossils of immature stages. It is well written in clear English, though the parts of the structure of the descriptions is overly repetitive, see below. The manuscript makes good use of the relevant literature. The figures are relevant and of good quality.

The descriptions are a bit cumbersome; I will suggest some simplifications to make them easier to read. Features of the head are described in terms of head segments (Line 239 onwards), yet the only visible manifestations of these are the mouthparts. Though head segmentation is an important aspect of basal insect evolution, it does not add any useful information in a description like this. I strongly suggest describing the mouthparts as such, no reference to the non-observable head segments to which they probably belong is necessary in this context.
Body segments are described one by one, but the descriptions are to a large extent identical (line 287 onwards). Here is is possible to just describe the general appearance of each segment, and then note the deviations from this pattern (if any) in individual segments. That will make the descriptions of the body a bit shorter and more readable without reducing the precision.
Experimental design: This point is of limited interest since this is not an experimental study. The technical aspects of the study, and the data analysis (in particular, the use of Dyar plots) are adequately described and discussed.
Validity of the findings: The systematic placement of the specimens is reasonable. I agree with the authors’ decision to place Dinobibio as Bibionomorpha, incerta sedis (but see comment on the decision to formally name it further down).

The authors have made a choice to formally name a species based on a larval specimen. Generally, it is not adviceable to name new species based on immature stages in insects since it is likely to lead to an inflation in the number of names, and this problem is especially relevant with fossil species since it is very unlikely that this specimen could ever be connected with certainty to an imago specimen, breeding and DNA comparisons not being possible. Most likely this species will stay «incerta sedis», the only likely way it could be associated with an adult insect is if a recent species with similar-looking larvae is discovered. On the other hand, it is also undesirable to have a large number of described but unnamed taxa in the literature. Trying to balance these concerns, I can agree with the authors that the least undesirable option in this case could be to formally name the species, in order to have a tag to attach to it. An alternative could be to use the approach the authors have chosen for the Mycetobia pupae, and describe it as an unnamed morphotype
Additional comments: Line 54: I think «destroying» is uneccessarily negative here, suggest «recycling» instead.
Line 66: Why «» around dystrophic? This is a standard term, delete unless there is reason to believe these lakes were in fact not dystrophic.
Line 116- «A full list of…» - move to line 99
Line 149 – Camera and what? Seems to be a word missing here
Line 183 – I believe that even monotypic genera need a generic diagnosis (though it will maybe be fairly identical to the species diagnosis). I think you have to give a short, generic diagnosis (Line 215), I suggest using those traits from the species diagnosis judged least likely to vary within a genus.
Line 224: «Acalypterate fly» is a fairly inprecise term. Is it possible to indentify further (to superfamily or family?).
Line 312: are there really «dozens» of them? Does not look so to me.
Line 321: should mention that the posterior spiracle is not notably enlarged compared to the others, as this sets this larvae apart from those known in Bibionidae.
Line 366: The body protuberances are quite erect in the specimen, this suggests they may have been more rigid than is the case with modern bibionid larvae.
Line 412 onwards: several of the species of Bibionomorphans described by Skartveit (2008) differ notably from recent counterparts and could possibly be the adult stage of this larvae. In particular, the three species placed in the genus Hesperinus and the enigmatic species Penthetria integroneura, which was tentatively placed in the genus Penthetria.
Line 432 onwards, description of Pachyneura sp. The contracted body suggests the specimen was desiccated before being encased in resin. This should be commented upon. As in the other descriptions, the descriptions of head and abdomen structures are unneccessarily long and repetitive. The same goes for the Mycetobia specimens.
It could be useful to briefly discuss the Mycetobia pupal morphotypes in terms of sexual dimorphism, which is often substantial in bibionomorph pupae. I would guess that morphotypes 1 and 3 are probably males and morphotype 2 probably a female, though this should be checked by comparison with recent material, if any is available.

Discussion
Line 1009: it is not difficult to erect new taxa, the difficulty lies in trying to relate them to other things afterwards.
Line 1076: body length is difficult to use to identify instars even in recent specimens – this should be døme by measuring rigid structures uch as head capsules.
Line 1149: «pristine» (meaning not affected by human activities) is not a very useful term for Eocene forests. I am not aware of any evidence of extensive logging back then.


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: FLY PALAEO-EVO-DEVO: IMMATURE STAGES OF BIBIONOMORPHAN DIPTERANS IN BALTIC AND BITTERFELD AMBER
Review round: 1
Reviewer: 2

Output:
Basic reporting: It is an interesting and well written manuscript, meeting the high standards of PeerJ. There are numerous minor errors throughout the text, including typos (e.g. “Mycetophylidae”, line 410), grammatical errors and double spaces, which should be corrected. All the manuscript would benefit from shortening. The literature is mostly relevant and properly cited (though several items are absent among the References, e.g. Azar et al. 2018, Paramonov & Salmela 2015, Wojton et al., 2018, 2019a, 2019b). The figures are of high quality, with appropriate legends.
Experimental design: The authors followed standard procedures of systematic paleoentomology. The fossil specimens are preserved in publicly accessible collections.
My only concern is giving a name to a new taxon based just on the larva and without a proper diagnosis. In the groups like Bibionomorpha, usually only male terminalia provide sufficiently distinct species-specific morphological characters. The probability of correct association of the named fossil larva with an adult male or female of the same species in the future is very low or even zero. The authors should ask themselves about pros and cons of such a controversial taxonomic act. Such a name will only feature in the lists of named fossil taxa, without any possibility to recognise the species (or genus) and further work with it. As a taxonomist (neontologist), I would definitely prefer not to give a formal name to a taxon where only larva is known. Interesting discussion related to the taxonomic descriptions without proper diagnosis may be found also in the paper by Jaschhof (2007, Paleontological Journal, 41: 103–106).
A further concern, related to the previous one, is that the immature stages of many genera of Bibionomorpha, especially outside Europe, are still unknown or insufficiently described. This applies also to Penthetria, mentioned as a possible relative to the new genus, where larvae are unknown in the vast majority of extant species. So there is a possibility that the larva described here as a new genus may actually belong to some described but insufficiently known extant genus or species. For me, this would be another reason why to avoid description of a new genus or species based on larva only.
Concerning the morphometric parameters and calculations based on fossil specimens, possible distortions of various body parts of the insect during the fossilisation process should be taken into account. This possibility is not commented on by the authors.
Validity of the findings: The new taxon may be valid but its practical identification and delimitation in the future will be difficult, see above. However, I see main value of this paper in the general overview of the diversity and abundance of some bibionomorphan immature stages in the Baltic and Bitterfeld ambers and in the contribution to the comparative morphology of dipteran larvae.
Additional comments: Immature stages of Bibionomorpha still represent a little known and challenging topic of study, both in extant forms and in the fossil record. Bibionomorpha really represents a megadiverse insect group but I cannot agree with the sentence of the authors in the Introduction that “Bibionomorpha includes numerous ecologically similar ingroups”. This sentence should be clarified or corrected. Even within a single family of Bibionomorpha, Cecidomyiidae, we can see various ecological adaptations of the larvae, including mycophagy, phytophagy, zoophagy, etc. In my opinion, the authors tend in various parts of the manuscript, especially in the Introduction, to overstatement and unjustified generalisation of the facts. In the Introduction they prefer to cite general textbooks instead of more specialised recent papers.
The exact phylogenetic position and interrelationships of particular subgroups of Bibionomorpha is still much disputed and challenging topic in current dipterology. However, the manuscript under review does not bring new insights to this debate. Actually, both Baltic and Bitterfeld ambers are apparently too young to be essential for the understanding of the origin and diversification of early bibionomorphan taxa. On the other hand, it is definitely useful to realise, in the context of the manuscript under review, that most of the taxa of Bibionomorpha found in the Eocene ambers represent extant genera or higher taxa.
Figure 1 deserves more explanation and improvements. It is actually not based only on the paper by Ševčík et al. (2016), as mentioned in the legend to this figure, because those authors did not include into their analyses taxa like Valeseguyidae or Rangomaramidae. Concerning the latter taxon, its concept and delimitation is especially controversial, so this name should not be used without further explanation.
The description of the new species is rather lengthy, giving the fact that many sentences are repeating. This section (and also several other similar parts of the manuscript) should be shortened. The sentence “The larva is clearly different from any modern representative of Bibionidae …” should be changed to “The larva is clearly different from any modern representative of Bibionidae, where immature stages are known...”.


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: ENVIRONMENTAL FATE OF THE ANTI-PARASITIC IVERMECTIN IN AN AQUATIC MICRO-ECOLOGICAL SYSTEM AFTER A SINGLE ORAL ADMINISTRATION
Review round: 1
Reviewer: 1

Output:
Basic reporting: No comment
Experimental design: No comment
Validity of the findings: No coomment
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: ENVIRONMENTAL FATE OF THE ANTI-PARASITIC IVERMECTIN IN AN AQUATIC MICRO-ECOLOGICAL SYSTEM AFTER A SINGLE ORAL ADMINISTRATION
Review round: 1
Reviewer: 2

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: As an effective anti-parasites drug, IVM has been widely used in the aquaculture industry. However, due to its hydrophobic property and high affinity to organic matter, the long-term accumulation of IVM in the aquaculture environment was recognized as the diffusion sources of pollutants affecting ecosystems. Finding a good bio-indicator to monitor the concentration of IVM in aquaculture environment will be benefit for subsequent operation to reduce the IVM residue level in environment. To this end, the authors in this manuscript established a system including an aquatic environment (water and sediment) and aquatic organisms (invertebrates, aquatic plants and fish) to mimic a real aquatic micro-ecological system. They measured the dynamic of IVM concentration in all different individuals included in this system after administrating IVM to fishes using gavage.  They found that there was a second accumulation peak in fish and invertebrates post administration 30 days for first time. They also found that the mud snail can be used as an indicator for the concentration of IVM in the whole aquatic system as the in vivo IVM amount in mud snails remained at a high level more than 70 days. Because the system they established could be more similar with nature ecological aquatic system, it is a most important part of this manuscript that is different from previous studies which evaluated the environmental residue of IVM. The authors should emphasize this point in discussion part and also need to point out the shortage or disadvantage of this mimic system they established to measure IVM residues in environment.
In general, the methods and results in this study are reliable and meaningful to the management of drug residue in aquaculture. However, some language issues in the manuscript let the paper difficult to read and understand. The manuscript would benefit from a critic review by the authors on English grammar and style issues.
In addition, some minor points and remarks have been addressed to shed light on the manuscript.
1.	IVM was found to accumulate in visceral mass twice, i.e. at 1 d and between 30 d and 45 d after administration, respectively. Why there was no second peak in muscle tissue? How to explain that?
2.	Secondary accumulation of IVM was found in organisms. Is it found at first time? Is there any reference to show similar discovery? Or is there any drug else showed the similar accumulation characters in fishes and other organisms?
3.	The “(Wall et al., 1987)” in line 64 should be deleted.
4.	The “minutes” in lines 115-132 should be replaced by “ min”
5.	The “ (Chen et al., 2007)” in line 246 also should be deleted
6.	line 254, “hour” should be “h”


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: CHARACTERIZATION OF DENGUE CASES AMONG PATIENTS WITH AN ACUTE ILLNESS, CENTRAL DEPARTMENT, PARAGUAY
Review round: 1
Reviewer: 1

Output:
Basic reporting: The manuscript is well written with appropriate context and table & figure. The authors have provided underlying data in very clear and properly organized format for any interested reader to re-analyze or integrate into another study.
Experimental design: The research questions are well defined and experiments and data collection are sound. The methods are described or cited from literature.
Validity of the findings: The data presented in this manuscript adds important information to the Dengue epidemiology knowledge. The authors have done a great job in organizing the data, applying statistics and presenting all examined parameters including the non-significant ones.
Additional comments: Though this manuscript is about Dengue in Paraguay, it would be informative to add in the introduction the presence and serotypes of dengue in and around the geographical site and study time. In the preceding years the Africa-Asia region had predominantly Dengue 2. A map in this context would be very helpful.
	Similarly, Dengue from other places have been reported more or less in the same format and parameters. Please add in the discussion how the symptoms associated with Dengue 1 found in this study match or differ compared to other Dengue serotypes and patient populations.
	Figure 3: Please reduce the number of asterisks representing statistical significance. Its clear that the traditional * < 0.05, ** < 0.01, *** < 0.001 does not necessarily add significance to the scientific comparison and depends on other statistical parameters like n. In the current set the data points from the two groups are mostly overlapping, and it would be really difficult to put a cut-off limit if they were really distinct.


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: CHARACTERIZATION OF DENGUE CASES AMONG PATIENTS WITH AN ACUTE ILLNESS, CENTRAL DEPARTMENT, PARAGUAY
Review round: 1
Reviewer: 2

Output:
Basic reporting: Clear and unambiguous, professional English used throughout. YES

Literature references, sufficient field background/context provided. YES

Professional article structure, figures, tables. Raw data shared. YES

Self-contained with relevant results to hypotheses. NO
Experimental design: Original primary research within Aims and Scope of the journal. YES (to Paraguay)

Research question well defined, relevant & meaningful. It is stated how research fills an identified knowledge gap. NO

Rigorous investigation performed to a high technical & ethical standard. YES

Methods described with sufficient detail & information to replicate.  YES
Validity of the findings: Impact and novelty not assessed. Negative/inconclusive results accepted. Meaningful replication encouraged where rationale & benefit to literature is clearly stated. YES

All underlying data have been provided; they are robust, statistically sound, & controlled. YES

Conclusions are well stated, linked to original research question & limited to supporting results. NO

Speculation is welcome, but should be identified as such. YES
Additional comments: This article is very important to Dengue studies in Paraguay, main because describe the last epidemic perfil in country in adults. But, I have some questions, listed below.

1. About number of patients. The authors think that with 231 patients (119 Dengue cases positives) could suggest just title viral and anti-DENV IgG positive as severity markers? Other factors could be included, such as comorbidity or pre-existing disease.

2. Many of the results are expected, aren't novelty. For example, little viral load to more days disease, after 6 day principally. I suggest having this information along with the results already expected and demonstrated in the literature, emphasizing that results corroborate what is already recognized.

Other suggestions to methods sections:

- Patient population and clinical samples.

Line 103 - 107: "Patients of all ages were enrolled from January to May 2018 if they presented with an acute illness (≤ 8 days) defined by 2 or more of the following: fever (measured or subjective), red eyes, rash, joint pain involving more than one joint, and/or diffuse muscle pain. Patients with fever and no other localizing signs or symptoms were included.

Reviewer: Could write as follow: "Patients of all ages were enrolled from January to May 2018 if they presented with an acute illness (≤ 8 days) defined by fever (measured or sujective) only or with 1 or more of the following: red eyes, rash, joint pain involving more than one joint, and/or diffuse muscle pain.

- Molecular detection.

Line 124: "Arboviral rRT-PCR testing was performed at IICS-UNA."

Reviewer:  "All rRT-PCR testing was performed at IICS-UNA" (If this information is right, if not, rewriter).


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: CHARACTERIZATION OF DENGUE CASES AMONG PATIENTS WITH AN ACUTE ILLNESS, CENTRAL DEPARTMENT, PARAGUAY
Review round: 2
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: This is a very well presented and informative manuscript.


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

Input:
Title: CHARACTERIZATION OF DENGUE CASES AMONG PATIENTS WITH AN ACUTE ILLNESS, CENTRAL DEPARTMENT, PARAGUAY
Review round: 2
Reviewer: 2

Output:
Basic reporting: The paper have clear and unambiguous, professional English used throughout, literature references, sufficient field background/context provided, professional article structure, figures, tables, raw data shared and self-contained with relevant results to hypotheses.
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The authors made some the requested modifications and justified the refusal of the others suggested.


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: COMBINED EFFECTS OF WATER TEMPERATURE, GRAZING SNAILS AND TERRESTRIAL HERBIVORES ON LEAF DECOMPOSITION IN URBAN STREAMS
Review round: 1
Reviewer: 1

Output:
Basic reporting: This paper reports an experiment examining leaf litter decomposition rates in mesocosms mimicking urban streams, at different levels of water temperature (10 vs. 18 ºC), grazing snails (presence vs. absence) and leaf damage (intact vs. damaged by terrestrial herbivores). The rationale behind this study is that (1) the above factors differ between non-urban and urban streams (the latter being warmer, with greater presence of grazing snails, and receiving more damaged litter), but (2) most available information about factors affecting litter decomposition comes from forest streams. While this is a valid argument and an interesting topic, the study presents important flaws, which I detail in the other sections below.

The text in general needs to be improved, and ideally checked by a native speaker. There are many cases where sentences are not concluded, or punctuation marks are not used properly. Verb tenses are often used incorrectly (e.g., L109, 114) and there are quite a few typos (e.g. “increasement”, “may be continue to increase”). Some paragraphs are too large, and consecutive paragraphs are sometimes poorly connected.
Experimental design: It is unclear whether the simulated warming responds to differences between non-urban and urban streams in the study area or to climate change. Some parts of the text seem to suggest that both impacts are addressed, while this is clearly not the case. So, first of all, the authors should be clear about what kind of impact they are studying (i.e., effect of urbanisation vs. effects of climate change on litter decomposition) and make it clear in the text. In fact, the temperature treatments used may or may not be realistic, depending on which impact they target. For example, while a difference of 8 ºC could easily occur between forest and urban streams, this difference would be too high if related to climate warming, based on projections for the end of this century, even under the worst scenario for China (a max. increase of 6.0 ºC, cited on L184).

Having said that, if effects of urbanisation are the main focus, then other issues arise. Urban streams differ from non-urban streams not only in terms of the factors manipulated in this study (i.e., temperature, invertebrates and litter inputs), but also, and very importantly, in terms of water quality and habitat complexity. Thus a proper experimental design comparing non-urban and urban streams should take these factors into account. This is obviously not the case here, as only one type of water was used (from an urban stream?) and habitat complexity was kept constant (no substrate). Moreover, if non-urban and urban streams are compared, but only grazing snails are taken into account, then the role of shredders in non-urban streams is underestimated.

The issues raised above suggest that the present design is more appropriate to test effects of climate warming than effects of urbanisation. However, if this is the case, it needs to be made clear, consistently, throughout the whole ms. Furthermore, the experimental treatments used need to be better justified, and more basic field information needs to be provided in order to support predictions. In relation to this, the hypotheses need refinement. For example, “increased temperature” should be used instead of “climate warming”. Any mention to leaf quality should be excluded, as it was not examined. Hypothesis 4 is not properly formulated; if synergistic effects are expected for any combination of factors, then this should be explained. In general, it should be made clear why hypotheses are novel and how they are based on previous evidence (ideally, providing references).
Validity of the findings: I wonder how damage by terrestrial herbivores actually altered leaf quality. By measuring N and P concentrations in intact and damaged leaves the authors would have been able to actually show that herbivory decreased leaf quality. Instead, they relied on the measurement of “specific leaf weight”, which only indicates a relationship between leaf mass and area. While this leaf trait is often related to leaf toughness and a good indicator of leaf palatability, at least for shredders, I’m not sure how this applies to leaves that have been grazed by terrestrial insects. What I mean is, if leaves present holes due to insect grazing, then the relationship between leaf mass and area is clearly affected, but that does not necessarily mean that the remaining leaf material is of worse quality than that of an intact leaf. Pictures of intact vs. damaged leaves might be helpful, but only pictures of leaves at the end of the experiments are provided.

The data analyses section is quite confusing. In order to improve clarity I suggest to split it into different paragraphs and, importantly, clarify which analyses were used to test each of the hypotheses. For example, it is unclear why two different measures of litter breakdown rate (k and % litter mass loss) were used; this should be justified, otherwise choose one of them and discard the other. It is also unclear why temperature sensitivity was examined. Lastly, the explanation about what additive and synergistic effects are seems superfluous.
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: COMBINED EFFECTS OF WATER TEMPERATURE, GRAZING SNAILS AND TERRESTRIAL HERBIVORES ON LEAF DECOMPOSITION IN URBAN STREAMS
Review round: 1
Reviewer: 2

Output:
Basic reporting: The study performed by Xiang and colleagues is exciting and provide many relevant informations regarding the impact of urban streams on a key ecosystem process, litter breakdown. However, my main concern is with multiple topics of data analysis, which compromises the quality of the work so far. On the other hand, the authors can overcome these issues by performing more adequate analysis and text reorganization. Also, the presentation of results (including figures and tables) can be improved by the synthesizing results description and prioritization of the core of the study (i.e., litter breakdown). I have provided several suggestions to improve this important work in the sections below.
Experimental design: Material and Methods: the ‘leaf litter collection’ section should be first than ‘experimental design’.

Lines 168-169: I never seem before this kind of methodology to estimate growth rate, i.e., blotting dry alive individuals before and after the experiment. You provide some references to backup this procedure or at least justify why common procedures such as size-to-body mass curve estimations (i.e., measures of size to estimate body dry mass of individuals; Smock 1980, https://doi.org/10.1111/j.1365-2427.1980.tb01211.x) were not used.
Also, explain in details how growth rate was calculated.

Line 172: describe mesocosms here and not on line 136 and also provide more details about mesocosms installation and maintenance. Photos and schemes of mesocosms distribution within the study site would be very helpful and illustrative (even in grey scale or at least in the supplementary material).
Line 175-176: 10.00 ± 0.01 g instead.
Line 176: replace ‘unconditioned’ by ‘dry’.

Data analysis
The data analysis section should report only the analytical procedures related to the exploration of your data and/or hypothesis testing but not formulas or any other calculation to obtain the final response variables; otherwise, this section would be too messy. Thus, I suggest you move the breakdown rate estimation, growth rates, snail tissue dry mass (note that it was the first mention!), etc to their respective sections above.
Please, use different paragraphs for different subjects such as main tests (those testing the study hypotheses), secondary tests (testing any assumption, i.e., ‘specific leaf weight differences between intact and damage leaves’) and other estimations (i.e., synergies among predictors).
Regarding the analyses performed, a Student’s t-test is not adequate to time-series data such as water temperature because it assumes independence of measures, which is not your case – i.e., as you recorded one value of temperature at each hour for the same mesocosms. An adequate analysis, for instance, is a repeated measure ANOVA or a linear model using a temporal correlation component (Quinn & Keogh 2002, Zuur et al. 2009)

Quinn, G. P., & Keough, M. J. (2002). Experimental design and data analysis for biologists. Cambridge University Press.
Zuur, A., Ieno, E. N., Walker, N., Saveliev, A. A., & Smith, G. M. (2009). Mixed effects models and extensions in ecology with R. Springer Science & Business Media.

Regarding the analyses of water parameters, first, I did not understand why they were conducted because you have not stated any hypothesis for it (i.e., there were not previous expectation to justify such analyses). Also, repeating several times the same analysis for different parameters (in this case, water parameters) is not wrong, but increase the probability of Type I statistical error. Additionally, most of those water parameters should be very correlated (DO in % and in mg/L; conductivity and turbidity and so on) and then would be difficult (or even impossible) to determine if a parameter have changed as a function of a treatment or conduced by any other parameter.
Litter mass loss and breakdown rates are intrinsically correlated as both use the same set of data with slightly differences in their estimates. Thus, there is no need to use both in data analysis. I suggest you maintain litter breakdown (and only report the % of litter mass loss in the results, excluding it from figures and tables also) to be consistent with the estimation of k-total, k-microbial and k-snails, which are very interesting and relevant to unravel some of the effects being tested here.
Regarding snail growth, the authors should test if there was actually any (statistical) difference between initial and final mass of snails (that is, if there was growth). Looking for Figure 5a (especially for standard errors) it seems there is no growth (i.e., higher variability that surpass the zero line).
Finally, a much more serious concern (than normality) for any kind of parametric analysis is the assumption of homogeneity of variances among the levels of a treatment (see Zuur et al. 2009, Chapter 2, for a deeper discussion). Transforming data is not always a guaranty to the adequacy of this assumption. I suggest you present boxplots using litter breakdown rates (as your main response variable) in relation to each treatment, as a supplementary material – as visual inspection of this assumption is a much more recommended approach than any other hypothesis testing approach.
Validity of the findings: Results
Water parameters: should be described in the methods, as they are description of conditions experienced at each treatment (and not related to any hypothesis being tested directly, as I pointed above). Also, it is obvious that temperature would affect parameters such as pH, conductivity, dissolved oxygen and turbidity, as all are temperature dependent; please, make this clear.

Specific leaf weight: should also be described in the methods, because this comparison ensures you indeed have used leaves with different ‘qualities’ in your experimental set-up.

Leaf litter decomposition: Importantly, when a significant interaction (e.g., temperature and snail presence) is detected it means you cannot interpret each individual factor per se, because they are dependent of each one. In this case you should only report the interaction and discuss it; please, review it throughout the text.
Would be more interesting and informative if you report averages ± SE for each level of a treatment (instead of a value for the whole treatment as you did in line 273), when a difference was detected.
Indicate the % or proportion of difference when a lower or higher rate was reported. For instance, “Microbe-mediated litter breakdown rates were also XX% lower than damage leaves …” (lines 288-290).

Lines 268, 270, 272 and so on: Do not use decimal places when presenting %, as will be easy to read.

Discussion
Warming enhanced litter decomposition: the authors could provide a better contextualization of this effect in urban streams (mainly comparing the present results with others in similar conditions or regions) and try to put their findings into a real world context.
Please remove the paragraph between lines 332-346 as it do not add anything novel or contribute to your work.
The presence of snails accelerated litter decomposition: Lines 349-354: It is a repetition of information already present in the introduction; please start this sub-section on line 355.
Line 355: add “where shredders are often scarce or absent” after ‘urban streams’ between commas.
Line 363: replace ‘normal’ by ‘sufficient’
Lines 371-373: this idea should be further developed, as it is one of the most interesting at all. In what conditions and land uses should be expected the greater changes?

Terrestrial insect herbivores retarded litter decomposition: The authors should pay special attention to the fact that while specific leaf weight can be negatively correlated with nutrient concentrations, it is also is with secondary compounds. It means leaves are one of two options: better defended physically (higher toughness) or chemically (higher content of toxic compounds), but not both (Boyero et al. 2007, https://doi.org/10.1038/s41598-017-10640-3)

The interaction of water temperature, snail and litter quality on litter decomposition: I found this section too speculative and with many low-relevance arguments, especially because this combined influence of three factors is of most relevance of disturbed streams, such as the urban ones. The authors could explore much better the idea of higher metabolic demands of snails and its implications. Also, the snail mediated litter breakdown indicates that intact leaves are more prone to temperature effects than damage leaves; the microbial mediated litter breakdown evidence the temperature effect on microbial litter processing (independent of leaf type) and give some clues about why snail litter breakdown are intensified with temperature (i.e., higher microbial development and thus, higher nutritive quality of litter). This should be explored further.
Additional comments: Please double space all the text as will be easier to read.

The fourth hypothesis is quite uncommon; if the authors do not have an a priori expectation/hypothesis (a specific one) regarding interactions among factors then you should assume that this is exploratory rather than formulating a general and counter-productive hypothesis.

Line 16: I suggest you start the abstract with a broader context sentence such as the importance of litter decomposition and the prominent conversion of natural to urban streams worldwide; this will attract greater interest to your study.

Line 24: remove ‘winter’ as it is detailed in the methods and may give the wrong idea to the readers that your results may be only applicable to this season (but of course, further studies to test this assumption may be needed).

Lines 27, 28 and 30: round decimal places; use integers for simplicity.


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: COMBINED EFFECTS OF WATER TEMPERATURE, GRAZING SNAILS AND TERRESTRIAL HERBIVORES ON LEAF DECOMPOSITION IN URBAN STREAMS
Review round: 2
Reviewer: 1

Output:
Basic reporting: The authors have addressed my previous concerns and have significantly improved the writing. I have attached a document with further editing and a couple of minor comments.
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: COMBINED EFFECTS OF WATER TEMPERATURE, GRAZING SNAILS AND TERRESTRIAL HERBIVORES ON LEAF DECOMPOSITION IN URBAN STREAMS
Review round: 2
Reviewer: 2

Output:
Basic reporting: The authors have done a good job in addressing my previous comments, however, there still are some issues which were not properly considered or further points that deserve attention (not raised before but mostly that appear only in this new version of the MS).
Experimental design: No comment
Validity of the findings: No comment
Additional comments: Lines 331-338 & 339-345: Should be moved to a subsection entitle ‘Litter breakdown’ before the data analysis part.

Lines 338-339: note that ‘litter mass loss’ was removed from the MS as the authors previously mentioned. Please, remove all mentions to this estimation throughout the text.

Lines 348-350: You should explain how Dataset 1 (which contains water temperature data) was aggregated to allow the authors to perform a t-test. The authors probably averaged temperature data over all the experimental period for each mesocosm, resulting in a total of ten values. This is acceptable, however, for diel temperature oscillations the same procedure were not performed – inferred based on the degrees of freedom of t-test on line 273 of the MS. To fix this issue, I suggest you first calculate daily temperature averages for each mesocosm, then calculate a measure of variability (standard deviation, variance or coefficient of variation over the experimental period) and finally test this variability using a t-test.

Lines 348-493: this information should go in the supplementary material, maybe in a section ‘Supplementary methods’ as the results of these analyses were previously reported above and are in support of the experimental setup but not a result/finding of the study.

Lines 524-525: this sentence is not correct, because you are referring to ‘treatments with snails’ but at the same time mention ‘total’, ‘microbial’ and ‘snail-mediated breakdown’, which all were estimated based on the difference between treatments with and without snails. Please, review it. For example, when you use total/microbial/snail breakdown rates you have to forget about treatments with and without snails and focus the comparison on warming and/or leaf damage occurrence.

Line 617: why did the authors use SLW in μg/mm2 instead of mg or g/cm2, which is a much more intuitive unit?

Figure 5. I suggest you delete the panel B (change of blotted dry mass), as this information is essentially the same as growth rate. Please, also remove any mention of change of blotted dry mass from the text for simplification.

Table 2. Initial litter mass loss should be delete, as indicated previously. Also, there is no need to log-transformed litter breakdown rates (to attain normality) as they are already very small numbers (< 0.1) – which suggest any transformation would not be efficient to this end but could remove important/interesting patterns in data. Thus, I suggest the authors to re-run this 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: NORRISANIMA MIOCAENA, A NEW GENERIC NAME AND REDESCRIPTION OF A STEM BALAENOPTEROID MYSTICETE (MAMMALIA, CETACEA) FROM THE MIOCENE OF CALIFORNIA
Review round: 1
Reviewer: 1

Output:
Basic reporting: The 3D model and CT scans are not presented in the supplemental material and cannot be reviewed.
Experimental design: See below.
Validity of the findings: See below.
Additional comments: Balaenopteridae is the largest family of living baleen whales. Its taxonomy is complicated due to morphological diversity and contradictions between the morphological and molecular data which are partly explained by introgressive gene flow (Árnason et al. 2018). 'Megaptera' miocaena is a well known, well preserved specimen which is, possibly, the earliest member of this family, and its re-description using the newest morphological and phylogenetic methods, as well as basic revision of balaenopterid taxonomy at the genus level, would be highly appreciated.

However, the work presented in the manuscript in its current form is far from being complete. The verbal description adds little to the original description by Kellogg (1922). The illustrations show only the surface scans lacking the photographs and the 3D component. The diagnostic traits are not specifically illustrated, and there is no comparative illustrations showing the closely related taxa. The illustrations of the periotic are especially non-informative, and the CT scanned inner structure is not presented, although reported in the Material and Methods section.

The differential diagnosis for the genus is not formulated that is the more important because there is no modern, updated diagnosis for genus Balaenoptera in the light of molecular genetic data. Moreover, while the genus Balaenoptera is pending revision, naming new balaenopterid genera will be debatable. The diagnosis in its present form is based on autopomorphies backing to phylogenetic studies by Marx and Fordyce (2015) and Slater et al. (2017), and no new phylogenetic research has been conducted here. Noteworthy, the apomorphies of the occipital shield are similar to the living Megaptera (lines 680-683), and the same can be said on the shape of the zygomatic process. The specific traits of the periotic bone also are similar either to Balaenoptera spp., or to Megaptera, as admitted by the authors. Thus, the diagnosis should be re-defined as a clear differential diagnosis at the genus level.

No comparison has been provided for 'Balaenoptera' ryani, which is the most closely related taxon for M. miocaena, according to the phylogeny by Marx and Fordyce (2015), and the comparison with B. bertae is rather limited. The comparative drawings showing these taxa are necessary, and taxonomic opinions on them will be helpful.

The comparative part of the manuscript, including the supplementrary material, seems to be incomplete and needs to be finalized. In particular, this concerns the comparison with Incacujira (see the question marks and yellow highlights in the submitted version).

The taxonomic position of 'Megaptera' miocaena have been discussed in detail in two papers with the same conclusion.
As Deméré et al. (2005) reported, "this basal balaenopterid taxon is not a species of Megaptera and, pending additional study, should be placed in a new genus. ... Although originally assigned to Megaptera, the holotype does not possess any of the autapomorphies of the extant humpback whale..."
Marx and Fordyce (2015): "If the nesting of grey whales within crown balaenopterids turned out to be wrong, our analysis would still indicate that grey whales are more closely related to living rorquals than certain balaenopterid stem taxa (e.g. ‘B.’ ryani). Further evidence to support this view comes from the recent description of the bony labyrinth of a range of living and fossil mysticetes, which demonstrated the occurrence of a tympanal recess in eschrichtiids and crown balaenopterids, but not ‘Megaptera’ miocaena [69]. This complements our results, which consistently interpret ‘M.’ miocaena as basal relative to both living rorquals and grey whales (figures 2; electronic supplementary material, figures S2 and S3)."

Therefore, erecting a new genus here is a formal recognition of previous studies and opinions which have become a matter of general consensus among researchers and should be properly respected. At least, the comments by Tom Deméré and Felix Marx will be an important, if not necessary, part of the publication process.

The schematic phylogenies provided in the Figure 1 do not contain the references to the original sources.

Finally, naming a higher rank taxon of Pan Balaenopteroidea is not substantiated, as it is not clearly supported by phylogenetic study. On the contrary, there is a robust definition of Balaenopteroidea as the clade pooling together Balaenoptera, Eschrichtius and the genera which are closer related to them as to Caperea (Steeman, 2007; Marx and Fordyce, 2015), so adding a new rank instead of it is redundant.

Thus, the diagnosis, comparisons and corresponding illustrations should be reworked before the publication, and the previous works on phylogeny and nomenclature should be fully addressed.


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: NORRISANIMA MIOCAENA, A NEW GENERIC NAME AND REDESCRIPTION OF A STEM BALAENOPTEROID MYSTICETE (MAMMALIA, CETACEA) FROM THE MIOCENE OF CALIFORNIA
Review round: 1
Reviewer: 2

Output:
Basic reporting: The English is clear and easy to follow. I only made a few suggestions to slightly improve some sentences (constructions somewhat unusual, see annotated pdf).
The list of refences is sufficient, but the field background should be presented in a less ambiguous way, clearly stating the input of previous studies on our present knowledge of this taxon (e.g., geological age and phylogenetic relationships, see comments below and in the text).
The paper is well organized, and only a few short parts should be moved elsewhere (e.g., size estimates are provided in the introduction, but should be moved to the results).
All figures are fine and useful. However, see below my comments on the lack of some important views (lateral and posterior) and the probable complementarity between images taken from 3D models and photos. I also made suggestions in the text for the addition of labels in line drawings for diagnostic features.
Experimental design: This paper is about the review of a previously described extinct mysticete species. Part of the results can then be considered as new (description and comparison, with new illustrations), but as mentioned above I would suggest more clearly separating these new results from results taken from previous work (phylogeny, biostratigraphy). This should especially be clearly stated in the abstract and in figure captions.
This is a useful and carefully done revision, but apart from providing a new genus name, the main goals of the work could be more explicitly mentioned in the introduction and abstract.
Methods for the surface scanning are described in detail.
Validity of the findings: Morphological interpretations have been carefully undertaken. In a few places I question the interpretations, but I think all these minor issues should be easily fixed. As mentioned above and below, some anatomical traits could be better supported if additional views and photos (in addition the 3D views) were provided.
Most of the conclusions are well stated, but I think that the part on the divergence of crown Balaenopteroidea should be revised (see comments below and in annotated pdf). Again, this won't need much work, but will have consequences on the content of the abstract.
Additional comments: - Considering that the geological age of the type specimen and the updated phylogenetic relationships were already mentioned in previous works, it would be important to better highlight the main contributions of the present study to our understanding of this interesting taxon (see below the comment on constraints for divergence dates).

- The authors only provide views taken from 3D models as illustrations of the skull. In some cases, these views are undoubtedly better to identify morphological features (for example for ear bones). On the cranium, this is less obvious when looking at Kellogg’s original photos. I made comments in the text concerning regions that would deserve more detailed views, for example the vertex in dorsal view, with interpretive line drawing showing the relationships between nasal, frontal, parietal, and supraoccipital. Also, lateral and posterior views of the skull should be provided as main text figures. if not possible with the 3D model, then photos should be taken. Note that no posterior view was provided in Kellogg's original paper, so this would be especially useful.

- Line 714 : ‘Similarly, Norrisanima’s position outside of crown Balaenopteroidea…’ : This is not similar. For Pithanotaria the taxon is in the crown group, and thus indeed provides a minimum age for the divergence of the crown group. Here you deal with a stem balaenopteroid, so it does not provide a constraint for the crown group (Norrisanima may have existed well before the divergence of crown balaenopteroids, but also well after). I think that you only provide a constraint for the stem Balaenopteroidea. This section should be revised in detail.

- Please check for all taxa the way to provide authorship:
authors, date: for species in their originally described genus
(authors, date): for species that were later referred to another genus

- Figure 1: The caption should preferably be understandable without the main text, so I would suggest adding here at least one or two references for each topology figured here. Especially if no phylogenetic analysis was performed during this study.

- Please see the annotated pdf of the ms for additional comments and suggestions, both in the text and figures.

This is a fine and useful work, and even if the revision may take some time, it will certainly make a good contribution. I am looking forwards to seeing this paper published.

O Lambert


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: EFFECTS OF FERULIC ACID ESTERASE-PRODUCING LACTOBACILLUS FERMENTUM AND CELLULASE ADDITIVES ON THE FERMENTATION QUALITY AND MICROBIAL COMMUNITY OF ALFALFA SILAGE
Review round: 1
Reviewer: 1

Output:
Basic reporting: In most parts, the manuscript is clear and is well-structured
Experimental design: Reflection on the aim of study:
FAE producing inoculants are used to open up complex structure of plant fiber during ensiling to improve ruminal degradation of fiber. It is not common to apply them to provide substrates (with helps of fibrolytic enzymes) for a successful ensiling. This approach might actually have adverse effect on fiber digestibility in the rumen, something which could be speculated from the results obtained (see below)

Sampling:
Maturity stage of alfalfa should be mentioned. In addition, it is informative to provide data about size of the field and distance between the two sampling points.

Methods:
For DM determination, samples should be dried at 103°C. Drying at 65°C will not remove all the sample water. In addition, for silage samples, DM should be corrected for losses of volatiles during drying.

Statistical analysis:
The experiment comprises two factors: sample (sample 1 and sample 2) and treatment and therefore one-way ANOVA should not be used. Alternatively, I suggest to use mixed model and include sample as random factor. In addition, the method used for pairwise comparison should be mentioned.
Validity of the findings: Reflection on the results:
To evaluate the effect of treatments on DM loss during ensiling, I suggest estimating losses of DM during ensiling (i.e. DM in –DM out) instead of only presenting silage DM.

The NDF represents fiber fraction of the biomass.  The reduction of NDF during ensiling by LF+CE treatment suggests that this treatment would likely reduce ruminal degradation of fiber. This is because easily degradable parts of the fiber, which are otherwise utilized by rumen microbes, are already degraded and utilized during ensiling, increasing the concentration of low degradable parts of the fiber. In order to confirm /reject this theory, silage samples can be subjected to in vitro ruminal degradation assays.
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: EFFECTS OF FERULIC ACID ESTERASE-PRODUCING LACTOBACILLUS FERMENTUM AND CELLULASE ADDITIVES ON THE FERMENTATION QUALITY AND MICROBIAL COMMUNITY OF ALFALFA SILAGE
Review round: 1
Reviewer: 2

Output:
Basic reporting: This article is well written in English. However, sufficent introduction and background should be include to demonstrate. For example, manyprevious experiments have been conducted to investigate the effects of FAE-producing LAB on silage quality and animal performance, and you should introduce recent findings, main opinions and shorting points of FAE-producing inoculants, and explain why you selected L. fermentum as your silage additives.
   Authors should provide the raw data of bacterial diversity, espcially for high-throughput sequencing.Data on alpha diversity of bacterial community after 60 days of ensiling was not well/statistically shown in Table 5. The same question on relative abundance of certain genus such Lactobacillus, occurred in Figure 2.
   In addition, a hypothesis is necessary for your introduction.
Experimental design: more details were for morphological, physiological, and biochemical characteristics of L. fermentum17SD-2. In addition more details for experimental design and samplings will help our understanding. Please note the true replicate in experimental design, especially for microbial diversity analysis.
Validity of the findings: no comment
Additional comments: This research is helpful to develop effective LAB inoculants to enhance fermentation of high fiber-contained silage. However, authors will need to revise the manuscript, if published in Peer J. The more concerning issues are as following:

ABSTRACT
More silage fermentation parameters such as pH, ammonia-N and volatile fatty acids were shown in abstract section.

INTRODUCTION
This article should include more relevant literature for sufficient introduction and background. Author should introduce the recent finds, main opinions and shorting points of FAE-producing inoculants, and explain why you selected L. fermentum as your silage additives. In addition, a hypothesis is necessary for your introduction.
L67-68, an opposite conclusion from their study was that inoculation of FAE-producing LAB did not alter the growth performance of finishing feedlot cattle.
L70, wrong reference，the inoculants used in the study of Chen et al (2017) was not FAE-produceing LAB.
L73, more references were necessary, such as Li et al (2019), Lynch et al (2014), Jin et al (2015), Addah et al (2014), etc.
L77, the objectives of this study is to (1) characterize FAE-producing L. fermentum (LF) and (2) investigate the effects of LF, celluase or their combination on silage fermentation and bacterial community?

MATERIALS AND METHODS
In this section, more details were for morphological, physiological, and biochemical characteristics of L. fermentum17SD-2. In addition more details for experimental design and samplings will help our understanding.
L88-89, explained why choose L. plantarum TS15.2 not other no FAE-producing L. fermentum as negative control.
L90, the diameters of a clear zone was ?
L91, delete the reference of Xu et al. (2017)
L100, why not add growth temperature of <20℃. If not, the description of wide growth range is not appropriate.
L106-107, how did you gain the two DM contents, by wilting or advanced maturity stage? And, each sample as group was randomly?
L117-119, more details for experiment design. Provide true replicates.
L149, samples for microbial diversity analysis was from the three replicates with some treatment?
L161, is it bacterial diversity analysis?
L175, data on alpha diversity of bacterial community after 60 days of ensiling was not well/statistically shown in Table 5. The same question on relative abundance of certain genus such Lactobacillus, occurred in Figure 2.

RESULTS AND DISCUSSION
It is well known that alfalfa is not easily ensiled for higher buffer capacity and relatively lower WSC content as compared with grass. In fact, LAB dominated microbial population in control silage. Meanwhile, the count of yeasts was below 5.0 log10 cfu/g FM. Why author concluded that poor fermentation resulted from low LAB counts, but not from high buffer capacity? Author should focus on the CP contents between treatments. Importantly, author should explain why LF+GE increased richness in silages from sample 1, but decreased that from sample 2. Moreover, why the bacterial diversity of ensiled alfalfa was higher than that of fresh alfalfa? Importantly, explain how chemical compositions such as WSC, DM, NDF and NDF affected Lactobacillus, Enterococcus and Weissella.

Others
Aata on alpha diversity of bacterial community after 60 days of ensiling was not well/statistically shown in Table 5. The same question on relative abundance of certain microorganism such Lactobacillus, occurred in Figure 2. The relative abundance of top 10 genus should be included, and that of certain microorganism such Lactobacillus should be statistically illustrated in Figure 2 C and D. If possible, please change the Figure A and B as PCoA.


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: EFFECTS OF FERULIC ACID ESTERASE-PRODUCING LACTOBACILLUS FERMENTUM AND CELLULASE ADDITIVES ON THE FERMENTATION QUALITY AND MICROBIAL COMMUNITY OF ALFALFA SILAGE
Review round: 2
Reviewer: 1

Output:
Basic reporting: Line 99-105: Explain how the evaluations were performed: (e.g. how did you measure salt tolerance or how did you measure bacterial growth?)

Line 123: The sentence “Each sample as group was randomly” is linguistically wrong.

Line 148: Did you correct silage DM for losses of volatile during drying?

Line 148: The sentence “DM contents and ground to pass a 1.0 mm screen for chemical analysis” is linguistically wrong.

Line 155-159: Why did you centrifuge the bacterial suspension twice?

Line 199: Replace “indigenous” with “epiphytic”

Line 305 and 350: Degradation of fibre during ensiling is not necessarily a good thing. See pubmed: 11063325

Figure 1: Stated in the method and material that bacterial growth was measured by measuring OD.

Table 3, 4 and 5: All the abbreviations should be defined. In addition, the values of main factors together with SEM should be shown.
Experimental design: No comments
Validity of the findings: The study provides useful information about the effect of feruloyl esterase producing inoculant and cellulase and their combination on fermentation quality of alfalfa silage. Unfortunately, the author did not provide information as to how the treatments affected the digestibility of silage. It is important to consider that improving fermentation quality of silage at the expense of silage digestibility is not desirable.

The author replied in the rebuttal that in vitro evaluation of silage digestibility was not in the scope of the study.  However, without this information the paper is incomplete in my view. Therefore, as it is I cannot recommend this article for publication.
Additional comments: No additional comments


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

Input:
Title: EFFECTS OF FERULIC ACID ESTERASE-PRODUCING LACTOBACILLUS FERMENTUM AND CELLULASE ADDITIVES ON THE FERMENTATION QUALITY AND MICROBIAL COMMUNITY OF ALFALFA SILAGE
Review round: 2
Reviewer: 2

Output:
Basic reporting: No comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The manusctipt "Effects of ferulic acid esterase-producing Lactobacillus fermentum and cellulase   additives on the fermentation quality and microbial community of alfalfa silage" (#35821) has been revised well.


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: EXOGENOUS APPLICATION OF NITRIC OXIDE DONORS REGULATES SHORT-TERM FLOODING STRESS IN SOYBEAN
Review round: 1
Reviewer: 1

Output:
Basic reporting: 1. Clear and unambiguous, professional English used throughout.
2. Introduction needs simplification.
3.	Line 58, 59 “Over time, submergence……plant death” and “High ethylene ……root elongation” is lacking in excess.
4. Literature references, sufficient field background/context provided. But please modify the format of the references in the text, for example, line 46,55, 129, 131, 355, 444 , and check and revise  the references section of the article.
5. What does the different uppercase and lowercase letters represent in Figures? Please add.
Experimental design: 6.	 The title of 2. 1. does not correspond to the content, and this section does not describe the flooding stress.
7.	Please add water, fertilizer, light, temperature and humidity management to the seedlings planted behind plastic pallets in part 2.1.
8.	On line 154- what is stage VI? On line 159 what is stage 3 (V3)?
Validity of the findings: 9.	No references should be included in the results .
10.	Line 450 "Recently, very little information are available on the ameliorative role of NO under flooding stress" has nothing to do with this study and is suggested to be deleted.
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: EXOGENOUS APPLICATION OF NITRIC OXIDE DONORS REGULATES SHORT-TERM FLOODING STRESS IN SOYBEAN
Review round: 1
Reviewer: 2

Output:
Basic reporting: In this manuscript, Khan et al., report the effects of NO donors on various parameters (SNO groups, gluthatione activity, superoxide, ABA content, mRNA transcripts of NO associated genes) in waterlogged soybean cultivars. While it is clear that NO plays an important role in abiotic stress signaling and more robust data on NO dynamics in flooded plants is welcome, it is partly because NO applications and dynamics are hard to study and interpret. While I am happy with the readability of the manuscript and the authors attempting to contribute data to the field, I have several issues with the current manuscript with regards to reporting and experimental design that in my opinion require clarification. I will also raise some minor comments.

English writing/grammar: Report is relatively clearly written and structured but should be improved on English grammar/vocabulary. Some English could be improved such as in the first sentence of the introduction (editor or English native/expert should go through the manuscript). 45: FloodING is…. And the first sentence of the conclusion: 450: Recently, very little information IS… (and it is not recently that little information is available).
Also, punctuation and lack of use of capitols is sometimes wrong. I.e. 101: Therefore, etc..

Intro and background: Flooding and soybean losses are well reported and are adequately referenced. Also the overview of NO production and interactions with various pathways are well described. However I have issues with the one sided use of NO as a signaling molecule that is solely reported to be beneficial for abiotic stress tolerance. There are many studies described where NO actually is negatively correlated with survival for instance in the context of the N-end rule pathway or phytoglobins (Gibbs et al., 2014, Hebelstrup et al., 2012, overview in Sasidharan et al., 2018). This should be addressed in the introduction. Additionally, the application of NO (donors) is not necessarily adaptive itself, but induces pathways such as ethylene or cyanide induced signaling which are directly adaptive (Mira et al,. 2016).

Figures are described and labeled adequately, although sample size should be clarified. In the raw data, it appears only 3 replicates were used for all experiments. But from the methods sections these appear to be the means from each separate experiment consisting of 5 replicates (itself containing 9 pseudo-replicates?). Not really clear at first view. Raw data is only the mean processed values of each experiment. Aren't these relative (for instance for RT-qPCR results, raw CTs, technical replicates, housekeeping gene results) How were these calculated? What are the units?
Experimental design: The biggest issue I have with the r is the experimental design that is not clear enough to me to interpret the results (and the meaning of these results).

First of all, the soybean plants are waterlogged in either normal water or supplied with an NO donor. The authors should describe how this NO donor is applied, is it in the water that is used for waterlogging? When? At the start or at the 3h and 6h time-points? SNP and SNAP donate NO GAS (only in the light) and this will thus penetrate and affect only above water tissues (gas diffusion underwater is reduced 10.000 fold). In addition, SNP and SNAP action decrease rapidly in the light and produce important side effects such as cyanide production . Sole cyanide application effects have mimicked the effects of SNP as an NO donor, thus attributing the effects to KCN application and not NO (Wodala et al., 2010). Therefore, the application should be very well described and the results should be very carefully interpreted and in this instance can’t necessarily be attributed to NO alone.

Secondly, since waterlogging is used as a treatment, what are the harvested tissues? Leaves, whole shoots? Of the above water or submerged parts of the plant? This is very important to clarify, as the submerged parts will have a drastically altered physiological state compared to the above water parts.
Submergence is associated with hypoxia stress, but this will not be the case for the above water tissues (see any plant flooding review), and possibly not even for the below photosynthetic parts in the light. The tissues affected possibly by hypoxia, ethylene, ROS/NO, CO2 accumulation are the submerged parts, but the treated parts with NO donors are predominantly the above water parts (see first issue). In my opinion, the authors should state very clearly what and why this experimental design was used before anyone can draw any conclusions on the obtained results.

Finally, what are the positive controls to show that the NO donor treatments actually increased NO levels? If it is not possible to measure NO levels directly, could an NO scavenger be used simultaneously to attribute the discovered results to actual NO effects, and no for instance KCN byproducts?

This does not mean that the obtained results are not useful for the research field, but should be placed into context of the experimental design which is impossible for any critical reader right now. I would gladly see any revised version where the experimental design is clarified.
Validity of the findings: See above, as the validity of the findings is related to experimental design.
Additional comments: -