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: BIOGEOGRAPHY, DIVERSITY AND ENVIRONMENTAL RELATIONSHIPS OF SHELF AND DEEP-SEA BENTHIC AMPHIPODA AROUND ICELAND
Review round: 1
Reviewer: 1

Output:
Basic reporting: The authors present an interesting and comprehensive dataset of peracarid crustaceans off Iceland covering an extended bathymetric range from continental shelf to abyssal depths. The analysis is based on high taxonomic resolution which is rare for any such macroecological work and therefor of great value. The English language use is very good, the manuscript is well structured and easy to follow, and the figures are clear and follow the methodological description.  I do endorse publication of this work following some revisions, the nature of which I have described further below. I congratulate the authors on this interesting piece and am looking forward to reading a thoroughly revised version, once published.
Experimental design: The methodological approach is sound and well justified.
Validity of the findings: The findings are useful and valid within a local as well as global marine biodiversity context.
Additional comments: Abstract and throughout manuscript:
Define better what you mean by climate change effects on the area under investigation, and how these effects may alter/shift diversity and assemblage distribution across the 4 zones (conclusions/outlook).

Can the 4 clusters/zones be characterised by predominant peracarid families/genera or similar? In traditional assemblage analysis, it is common to name species assemblages by for example choosing numerically abundant taxa to name these. These could represent taxonomic state, ecology (eg feeding or specific reproductive mode) that help the reader associating an association with habitat type or ecological niches, and similar.

Why is there no overall representation of diversity – depth pattern irrespective of the 4 zones? I would suggest plotting this for the overall dataset, which then will be eluded upon by discussing the environmental variables for each zone and attempting to identifying the factors that may govern the observed overall pattern the most (similar to what is done for the 4 zones in the present manuscript). Also, this will allow the authors to compare overall diversity -depth pattern with those from other parts of the ocean, in particular for the North Atlantic (eg. Carney, R. S. (2005). Zonation of deep biota on continental margins. Oceanography and Marine Biology: An Annual Review 43, 211–278.), and whether the Icelandic patterns conform to other observations and given that eg temperature changes with depth are less pronounced than at lower latitudes. Incorporate this aspect throughout the manuscript, please. I believe this – though generalising aspect- makes the paper more appealing to macroecologist/physiologists and a wider audience rather than those only interested in peracarids and ‘Icelandic’ diversity.

Results section:
The environmental variables characterising the 4 zones should be better defined/described; something like “low temperature” or “higher pH and dissolved oxygen” is too cryptic. Prvode means …ranges and similar and supporting references.

Discussion:
Effects of depth and hydrostatic pressure. The two citations provided are valid, but the authors may wish to add to the discussion by further referring to a review in Biol. Rev. (see below) that helps identifying physiological factors that govern depth diversity patterns. Depth alone has traditionally been used as a variable, but it indeed is hydrostatic pressure rather than depth as a location proxy that determines diversity patterns. The text from line 351 on will benefit from this discussion of pattern-determining diversity with depth/increase in hydrostatic pressure.
Please do consider the following review within this context:
Brown, A., S. Thatje (2014). Explaining bathymetric diversity patterns in marine benthic invertebrates and demersal fishes: physiological contributions to adaptation of life at depth. Biological Reviews, 89: 406–426.

Line 499: split “webelieve”


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: BIOGEOGRAPHY, DIVERSITY AND ENVIRONMENTAL RELATIONSHIPS OF SHELF AND DEEP-SEA BENTHIC AMPHIPODA AROUND ICELAND
Review round: 1
Reviewer: 2

Output:
Basic reporting: The authors clearly stated the purpose and problems to solve in the manuscript. The title reflects the contents of the paper. The abstract describes the vital information in the work, and the introductory section sufficiently explains the framework and problems of the research. However, I found some obvious errors, and I have made several comments on the abstract and introduction sections as below:
Line 49. How many records? The dataset includes genus-level data. Thus, I think it is not 357 species.
Line 53. Why only salinity and temperature? How about depth? As the authors mentioned that the most important environmental variables were temperature, depth, and salinity (line 276-278).
Line 55. Is it GIF or GIFR?
Line 65. The same word was used in one sentence. Suggest revising.
Line 96. Please check the spelling for the cited literature.
Line 121-124. Missing citation.
Line 128. Can be more specific, which literature?
Line 129. How many records?
Experimental design: This is a well-defined research question. However, I have some notes on the methods section as below:
iNEXT package user guide (page 2 & 5) has shown that the package allows incidence data to perform analyses if abundance data is not available or incomplete. The authors indicated that they did not have complete data on abundance (line 143-145). Please explain why the authors used abundance data.
Line 139. How many occurrences? Where is abundance data?
The dataset includes genus-level data; thus, it needs to be clarified how many species and genus.
Line 140-141. This sentence can be omitted as a bit confusing. It will be more concise if the authors refer to a table that contains such information.
Line 141. Can be more detail on “extensive literature search.”
Line 143. Please be more specific, how many?
Line 186. I could not find Table or Figure S3.
Check the requirement for the first mention for an acronym.
Line 198. Check the requirement for the first mention for an acronym.
Line 217. Can be more specific, what is being compared? Which literature? What information?
Line 218. Check the requirement for the first mention for an acronym.
Validity of the findings: The data seem robust, although there are some points that need to be clarified as below:
Table 1. Is it Vel or Velo? Please be consistent.
Table 2. Suggest revising Nr. to No and check the writings in the “species” column.
Some genus levels are still found in this table and were not given a numerical code as indicated by the authors in line 245.
Figure 1. Why are some lines in hexagons bold?
Figure 2. I could not find the appendix file that the author refers to in the figure.
Suggest removing vertical writings on the x-axes as legend has been provided
Figure 3. Missing legend, which one is Fig. 3A and B?
Figure 4. The Shannon-wiener index value increases with biodiversity. Conversely, for the Simpson index, a higher value indicating high dominance or low biodiversity. Please explain why in Fig. 4C coastal cluster has the highest Simpson value.
Supplemental tables. I would suggest combining those two files into a table and add information on the abundance data dan reference. The dataset is not massive, and I think it is worth moving it to the main section.
Line 244. How many occurrences data?
Line 245. There is no numerical code after the genus name in the dataset, such as in row 19. It only written Ampelisca Krøyer, 1842.
Line 271-289. How about other environmental parameters were shown in Fig. 3? Why no mention of other environmental parameters such as phybio and velo?
Line 318. How many individuals?
Line 330. The authors’ claim that the study was built on an extensive data set is a little bit excessive considering that the authors did not provide complete data on the occurrences and abundance and only employed 357 amphipods (species and genus).
Line 334-336. Why is the sentence about comparing diversity in the Nordic basins and the deep-sea coming after the authors’ statement that GIFR had a strong overlap with a coastal assemblage? Suggest adding a sentence that explains the cause of the overlap.
Line 364. Suggest revising the “incl.”
Line 403-414. This paragraph contains unfinished sentence, such as in line 406 and the extensive discussion part almost obscuring the main messages.
Line 416-500. The main point that I get from the “Diversity trends” section is that the diversity of the shallow clusters was higher than the deep clusters, which is valid considering the results presented in the manuscript. However, it could be written in a much shorter section. The section also needs to be organized to not obscure the main findings (idea) by supporting information.
Line 499. Missing space.
Line 518. Missing the article, please check other places too.
Additional comments: I think this is a relevant manuscript, but I feel that the discussion section should be simplified to make the main point clearer. Please complete the data and include all necessary information.


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: BIOGEOGRAPHY, DIVERSITY AND ENVIRONMENTAL RELATIONSHIPS OF SHELF AND DEEP-SEA BENTHIC AMPHIPODA AROUND ICELAND
Review round: 1
Reviewer: 3

Output:
Basic reporting: The manuscript is well written in clear unambiguous English, and clearly structured.  I have made a few minor suggestions to improve this on the attached PDF.  Literature is sufficiently cited throughout, although reference to the World Amphipoda Database is not cited despite being used and therefore needs to be included.  It is also necessary for the data tables and the dataset in OBIS to be cited in the paper.
Experimental design: The research aims are clear and relevant. The methods are well-described and in sufficient detail.
Validity of the findings: The findings of the research are interesting and clearly stated and discussed.
Additional comments: This is a well-written and clearly structured paper and should be published subject to the minor revisions 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: BIOGEOGRAPHY, DIVERSITY AND ENVIRONMENTAL RELATIONSHIPS OF SHELF AND DEEP-SEA BENTHIC AMPHIPODA AROUND ICELAND
Review round: 2
Reviewer: 1

Output:
Basic reporting: The manuscript deals with pattens of biogeography and diversity in a remarkable region. The manuscript is well written and constructed and should be published with minor revision.
The manuscript holds all of the relevant references. The background is clear and the questions asked are highly relevant.
I find the text on the older amphipod data could be extended. Much of the earlier data comes probably from the extensive Ingolf expedition, which was one of the major expeditions to the region at that time. Stephensen published a series of paper on the amphipods from the Ingolf expedition, but only a single of his paper is cited (but probably used in the analysis) dealing with material from the Ingolf. In a way this is a high-quality data. A fine mesh net to get small crustaceans was probably used for the first time in this expedition.
Figure 5 can be improved substantially. First, there are no labelling of A, B and C in the figure. The figure would benefit from having icons (dot, box, triangle, etc.) instead of differently colored dots (which are un-distinguishable on the print-out). The Deep South and the Deep North seem to differ in the left panel, but to be similar in the right panel. Distinguishing between the DS and DN would be beneficial in the right panel.
Line 86. Are the limits of the deep-sea at 150 m?
Lines 102-103. The ridge “… hinders direct flow …”. This is a strange statement.
Experimental design: The environmental layers should be presented more thoroughly. Are these layers near-bottom layers or shallow/midwater layers? Is the resolution within the layers similar in all parts of the region?
Also, some the layers should be termed somewhat differently. While temperature is without a doubt an environmental parameter felt by any species, I guess phytobiomass is hardly environmental parameter, but more of a community descriptor. But I feel that all of these should be used in the analysis.
The authors state that the mclust algorithm identified six clusters. It would be worthwhile showing these clusters in a figure, as it seems there is something interesting going on here, even though there are disproportionally large differences.
Validity of the findings: This is a solid work and very interesting results are coming out of the study. I can highly recommend its publication in PeerJ.
Additional comments: No additional comments


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

Input:
Title: THE COMPOSITION, GEOGRAPHY, BIOLOGY AND ASSEMBLY OF THE COASTAL FLORA OF THE CAPE FLORISTIC REGION
Review round: 1
Reviewer: 1

Output:
Basic reporting: The text is clearly and professionally written and meets all the major criteria of a good research paper, with clearly annotated and available raw-data. I've got no major comments, only some improvements to suggest below to polish off a very well presented study.

Some biogeographical terms being somewhat unclearly defined, especially from a non-specialist's perspective. I'd like to draw particular attention to "Afrotemperate" and "Afrotropical" (line 331, § Methods). These terms, while broadly intuitive, are potentially unclear as multiple geographical regions could be described as such. Does the Afrotemperate region, as far as this paper is concerned, refer to the somewhat more temperate southwestern corner of South Africa, or is it more synonymous with the pockets of Afromontane regions in the Cape, the Drakensberg and extending up towards eastern Africa? "Afrotropical" as used here could also be confusing, as this region is often considered to encompass all of Sub-Saharan Africa (when treated as one of the 8 or so global-scale biogeographic realms), and would then overlap entirely with any definition of the Afrotemperate/-montane region. Defining these terms more plainly, as has been done very helpfully for soil terminology (lines 370–373, § Methods), would make the text and the results themselves more apparent to non-specialist readers or readers from disciplines where these terms may have subtly different meanings.

The figures for this article are clear and insightful, but could be improved with slightly larger axis and label text, and some tweaks to the colour-schemes. In more detail, I have the following recommendations and comments (in descending order of importance):

1. Figure 6: I like this figure a lot, but it could be improved by making the range of circle-sizes greater (i.e., the biggest circle bigger, the smallest smaller), to make differences in the relative numbers of species more obvious. Ditto for the range of thicknesses in connecting lines. The raw species-counts for each circle could be annotated here for easy reference. Ditto for the no. shared species for the connecting lines. The colours of these circles don't add anything (though there's nothing with it, it is a bit distracting). Additionally, it would be useful to use the abbreviations for soil types used in this figure (CC, CS, SS, SH) in the main text as well, just to make the figure easier to use alongside the text.
2. Figures 4, 5: The data in these figures could be presented in a slightly more attention-grabbing way, perhaps. Maybe, seeing as it is mostly percentage/proportion data (4A–C, 5A,B), stacked-bars might simpler and make the data more self-evident? The colours in 4D and 5C are fine, but perhaps a different colour-scheme could be considered—maybe just grey and white? Also, throughout these figures in particular, the axis and labelling text could be bigger and thus more easy to read.
3. Figures 3, 7: Great figures—really creative and clear way to present these data! Although the colour helps to tell the squares apart, I did initially try to work-out what the colour represented for each family "as data". Perhaps, then, note in the caption that the colour is just for illustrative purposes in these figures, and does not represent any value or property of any of the families.
4. Figure 1: Elevation-colours could be more distinct from each other?

I feel these changes and improvements to the figures will help this paper really make its key findings and conclusions that much more apparent and stand-out.
Experimental design: The goal and knowledge was clearly defined. The authors set out to present a "sketch" of the history and features of this flora based on broad and comprehensive survey of its species, and they have done exactly that.

As mentioned above (see § Basic Reporting), the data are clearly annotated the collection process thereof is very well explained.

The data is notably supplemented by the authors' own observations, though the finer details of these observations are not presented. In principle this is not as transparent as it could be, but I doubt there are any problems with these observations, as this sort of data, along with expert-based data, is not unusual for this sort of study.
Validity of the findings: As mentioned above (see § Basic Reporting), the data are clearly annotated the collection process thereof is very well explained. Only fairly simple descriptive statistics were used in their analyses, and these are all perfectly sound.

The authors conclusions, following their insights and descriptions of the floristic data collated, are well substantiated and contextualised by previous literature and established ecological principles.
Additional comments: In its current manuscript-form, this article has a notably long discussion (22–23 pages) compared to the other sections (introduction: 4, study area and methods: 7, results: 7). While it is clear to me that the vast majority of the "conclusion-making" work and findings in this paper are made in the discussion, there are one or two parts of the discussions text that could go to the results section.

I have also made some smaller comments and suggestions on the text directly (see annotated review PDF).

To conclude, I think this article just needs a few adjustments for clarity's sake before being published, hence my recommendation for Minor Revisions. But, I must commend the authors nonetheless on a very impressive dataset and the conclusions they draw in this article.


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

Input:
Title: THE COMPOSITION, GEOGRAPHY, BIOLOGY AND ASSEMBLY OF THE COASTAL FLORA OF THE CAPE FLORISTIC REGION
Review round: 1
Reviewer: 2

Output:
Basic reporting: This is a well-written paper, sufficiently referenced, and very comprehensive. The language and style are mostly OK (see minor comments for some word change suggestions). There are no loops left hanging or unanswered.
Experimental design: Overall, excellently done. The study focuses on existing data (in various forms) rather than (it seems) on novel fieldwork or sampling any poorly known parts of the study area. There is no problem with this, and the bulk of the methods described are literature-based. As a small addition, the authors might want to add a line that no structured fieldwork was undertaken as part of this study, and I would suggest that details relating to any specific fieldwork that the authors did do be included (perhaps including a table of dates, no. of specimens collected, where / how identified, and where deposited).
Validity of the findings: The findings are detailed in much detail and very comprehensively. It is a positive contribution to understanding the Cape Flora and its origins, with useful implications for conservation.

Unless I missed it, a full flora /checklist would be a useful addition to the paper as supplementary data.
Additional comments: Minor comments:
l.50 - replace "huge" with another word
l.51 - delete "only
ll.57-59 - Suggest deleting "Note that" and starting with "Most biologists"; insert "as per"  after "limestone fynbos" and change citations to "Cowling & … (2001) and Rebelo  et al. (2006)".
ll.135-140 - very long sentence, best break up.
l.286 - perhaps standardize sub-tropical / tropical use?
l.324-325 - would there have been challenges assigning as sp. to more than 1 biome?
ll.431-440 - rather replace "woodies" with something less colloquial.
ll725-726 - citation for this statement?
l.903 - spelling of "autocthonus" - should be "autochthonous"
ll.1022-1052 - similar situation in the Hantam-Roggeveld, with long-term climate stability (winter rainfall) allowing for edaphic speciation on dolerites, sandstones and shales.
l.1089 - "in-situ" is this correct for this journal? italics? (and also for "sensu" through the doc - italics?)
l.1223 - "led" should be "lead"


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: EFFECT OF STORAGE CONDITIONS ON SARS-COV-2 RNA QUANTIFICATION IN WASTEWATER SOLIDS
Review round: 1
Reviewer: 1

Output:
Basic reporting: The manuscript is concise and well written. There are a few typos. Regarding the background/context I have two comments:

L56 More nuance is warranted here. A few of these studies used both the liquid and solids fraction of the influent together while another in Helsinki separated the two and then spiked. This is mentioned in the discussion but not here in the introduction. It would be useful to better describe the sample types used in previous studies.

L59 Per my reading, the study from Helsinki considered signal degradation in the solids fraction (separation by centrifugation) of influent samples.
Experimental design: This is a well designed and executed study and represents excellent work by the authors. The inclusion of MIQE checklist is also appreciated. One comment:

L137 The method of propagating error should be detailed further or incorporated by reference (maybe similar to Graham et al.?)
Validity of the findings: This is perhaps where I only have a real criticism/contribution to suggest:

L142 The justification for using a 68% confidence interval is compelling. But why not perform the analysis with both 68% and 95% intervals? Would all the significant differences remain? The authors should consider performing that analysis at the 95% confidence level and including those results in the SI. If the results are consistent, this would greatly strengthen the inferences. If they are not, then the authors can use their logic to justify their conclusions based on the type II error.

A few other comments pertinent to the discussion:

L228 Hokajärvi also examined solids (Table 2) that were pelleted via centrifugation and then spiked. It is worth discussing the current results relevant to their observations.

L243 This statement contradicts L59 unless the authors are making a distinction between solids separated by centrifugation and “settled solids”.

L244 Would the results of the current study be similar to those in Helsinki if 95% confidence interval was used instead of 68%?
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: EFFECT OF STORAGE CONDITIONS ON SARS-COV-2 RNA QUANTIFICATION IN WASTEWATER SOLIDS
Review round: 1
Reviewer: 2

Output:
Basic reporting: The paper is well written using clear professional English. A sufficient knowledge gap has been established. The article structure is good. Figures are of publication quality.
Experimental design: The experimental design is good and contains references.
Validity of the findings: No comments.
Additional comments: No Comments.


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

Input:
Title: EFFECT OF STORAGE CONDITIONS ON SARS-COV-2 RNA QUANTIFICATION IN WASTEWATER SOLIDS
Review round: 1
Reviewer: 3

Output:
Basic reporting: The language of the article is good. A few instances of things not being too clear or incorrect are pointed out below in specific comments. Some figures have misaligned graphs within them (see specific comment). Results section must be significantly reworked.

Here are some specific comments:

Lines 29-30: Is PMMoV a wastewater virus? Plant virus quasi-ubiquitous in sanitary sewage?

Line 36: Two of the 3 citations are by authors however several groups have outlined this, it may be good to cita a few more pieces of litterature: Kitamura (2021), Kocamemi (2020), D'Aoust (2021), Chik (2021). Balboa also published their preprint after you submitted (2021) it would be nice to acknowledge them during the review.

Line 48-49: "In such a scenario, samples are not stored, but are processed as soon as they are collected. Even if this is done as recommended, sample storage is essential." I think this paragraph should be reworked a bit. I can see what the authors are trying to do and justify why storage is important, but the result reads a bit clumsy. You could write that storage is important because of transport time, laboratories for analysis may not be located in the direct vicinity of the sampling sites (and so must be shipped), and bio-banks and reruns are sometimes necessary.

Results:

Lines 163-169, this sounds like methods... should this be in the methods to explain the experimental procedure? It seems like now this is a breakdown of the methods, but it's not results. This section should be moved to methods, and it's hard to textually understand what was done. Presenting the breakdown of when samples are testing in a table format would immensely help.

Line 185: ~50% An actual value would be very helpful here.

Figure 1: Would it be possible to provide more textually descriptive names for the samples. The sample names as they are are obtuse. Graphs are misaligned with eachother (particularly bottom 3), units for the bars are a bit bizzare and are not the same while the targets are the same, hence do not really provide easy comparison. The graphs should have the same X axises.

The whole section needs to be tidied up and tightened. Approximations of changes in concentrations is not really acceptable if the methods and numbers are to be quantitative. Needs rework. The information is valuable and the graphs (aside from the axises) are doing a decent job at dissimenating the information, however the textual description of the results section need to be brought up to a higher standard seen at other places in the manuscript. The language is too vague.

Discussion:

Lines 217-224: this is what should be in the results. This type of language, the n's, etc. The discussion section should describe of the implications of the observed results however, not give more results.
Experimental design: The materials and methods section was reasonably well done. The experiment was well planned, however the planning part of it is not well communicated to the reader. Furthermore, some inconsistencies from section to section exist making it appear as though multiple people worked on the sections but some formatting and writing style/choice differences are very apparent. The section must be reworked it make it flow better as currently from section to section its inconsistent.

Here are some specific comments:

Line 68: Settled solids from the primary clarifiers - are we talking of primary sludge? Settled solids may not be the optimal term for wastewater people.

General comment for methods - please provide information such as manufacturer and part numbers for equipment, supplies, reagents, etc. It's there in the digital PCR section of the methods but not other sections. It's making it evident different sections were written by different people, or that these details were not standardized throughout the document.

Line 71: Without going in great detail, adding one sentence briefly describing the method would help a bit for the readability of the methods. I feel stating high-throughput methods alone may not be descriptive enough. Ex; what makes the method high throughput?

Line 73: From the text alone the methods imply that we would have several points in between the ranges of days (7-122), however when the reader gets to later sections it is shown that the samples were tested at 7 AND 122, with nothing in between. Having a "-" with a range can  therefore be a bit misleading and actual experimental conditions should be stated, i.e. "stored at 4°C for 7, and 122 days). Furthermore, what was the rationale for the storage times selected?

Line 97: extra space before 500 copies and (.

Line 103: The information about being purchased from IDT could in my opinion be moved to Table 3. In addition, although it's pretty obvious for those of us in North America for international readers it may not be immediately apparent what/who IDT is. I think the company name (Integrated DNA Technologies) should be somewhere in the manuscript, either the first time you use the IDT acronym or as a footer for Table 3 explaining that probes and primers were purchased from IDT.

Lines 101-117: Whole section has different font for °C than the rest of the document. Please fix. Alt+248 (numpad) will do the correct degree symbol.

Some of the things that should have been in the material and methods section were found in the results section. This has been highlighted above in 1.

Discussion was significantly better written than the results section. Well done for that section!
Validity of the findings: Findings are probably valid but unfortunately with the results written the way they are currently it's not immediately possible for the reader to come to the same conclusions as the authors as the results are written as approximations (ie.: the
205 ratio of SARS-CoV-2 RNA to PMMoV RNA concentrations was ~50% lower in the treatment than the control. /// Differences between the experiments are shown in Table 3;
differences were between ~60% and 100%.). The section needs to be tightened significantly.
Additional comments: The manuscript was informative but the materials and methods and results sections need significant work. Some information that should be in M&M is in the results section. The results are given as vague approximations only.

The experimental protocol and outline of when samples were tested should be in a table format or in a format that's more easy for the reader to comprehend.

The discussion portion of the manuscript was significantly better quality than the materials and methods and the results. The manuscript is definitely worthy of publication as the information will be useful to the scientific community at large (and particularly those of us engaging in WBE) however those two sections must be reworked to bring them up to an acceptable standard.

Good job, and looking forward to seeing the final product published.


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

Input:
Title: EFFECT OF STORAGE CONDITIONS ON SARS-COV-2 RNA QUANTIFICATION IN WASTEWATER SOLIDS
Review round: 2
Reviewer: 1

Output:
Basic reporting: No comments. The manuscript is well written and ready for acceptance.
Experimental design: No further comments. I recommend for acceptance.
Validity of the findings: No further comments. I recommend for acceptance.
Additional comments: No further comments. I recommend for acceptance. Congratulations to the authors on well done study.


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

Input:
Title: EFFECT OF STORAGE CONDITIONS ON SARS-COV-2 RNA QUANTIFICATION IN WASTEWATER SOLIDS
Review round: 2
Reviewer: 2

Output:
Basic reporting: Changes made as requested. Very satisfied. Strong manuscript!
Experimental design: Changes done as required. Thank you!
Validity of the findings: Manuscript reads better. Thank you!
Additional comments: Great work, thank you for considering recommendations and applying them as required!

Few minor things to fix and you are good to go!

Line 240 of tracked changes doc: 107%, spacing between 107% and ±... to remain consistent with above.

Table 1, 1st column, formatting/spacing different for last cells (space in front)

Table 3, fonts and size of text differ significantly between cells... please standardize. (alternates randomly Calibri - Arial, and font size 10 - 8, for example)

Figures 1 and 2: Please ensure the "boxes" of the graphs are aligned. Please see my attachment where I outline some alignment issues. Nothing major but will improve look and feel of figures.

Please check the fonts of all captions for figures and tables - some are Arial, some are Roboto and some are Times New Roman.

Thank you!


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: LEARNING NEW WORD MEANINGS FROM STORY READING: THE BENEFIT OF IMMEDIATE TESTING
Review round: 1
Reviewer: 1

Output:
Basic reporting: This is a really neat, comprehensive and well-designed set of experiments that set out to address a current and important questions with clear (and broad) theoretical and practical value. Specifically, the authors examine adult participants’ ability to learn new words from stories than compared to an intention training paradigm. There were two key aims: (1) to compare performance in incidental and intentional learning paradigms, and (2) to examine the role of testing on longer-term retention. Adults learned new word meanings for familiar words better under intentional than incidental learning conditions, but there was also evidence in Exp 1 (and a trend in Exp 2) showing less forgetting for items learned incidentally (attributed to richer contexts supporting long term retention). There was very clear evidence of a testing effect in both intentional and incidental learning (and for both recall and meaning matching tests).
I have a few points about reporting of results in the Abstract:
(i)Regarding the description of results for Exp 1… “Results showed that intentional learning was more efficient”… what does “more efficient” mean and was this the case for both tasks? (ii)The abstract doesn’t connect Exp 1 and Exp 2/3. It would be useful to hint at why it’s interesting to remove the immediate test, particularly in the context of incidental learning (given testing takes it away from being so incidental). (iii) “Both cued recall and multiple-choice tests enhanced retention individually” It’s not clear what was done here. I.e., whether you examined whether immediate test performance contributes independent variance to test 2 performance, or whether you manipulated the presence of immediate recall versus immediate MC to show that both influenced test 2 as compared to having no immediate test. (iv)It would be useful to point out that the participants were adults in the abstract (given there’s a large associated literature on children).
Intro
Pg 7, lines 40-41. It’s highlighted here that incidental learning from natural linguistic environments (e.g., conversations, books, TV) is the main source of vocabulary learning, but references are taken from the developmental literature. These references might be more appropriate for supporting the idea that older children and adults learn most of their words through exposure to written texts: Nagy W., Herman P., & Anderson R. C. (1985). Learning words from context. Reading Research Quarterly, 20(2), 233–253 and Sternberg R. J. (1987). Most vocabulary is learned from context. In McKeown M. G., & Curtis M. E. (Eds.), The nature of vocabulary acquisition (pp. 89–106). Hillsdale, NJ: Erlbaum. You also cite the Batterink and Neville (2011) paper later on, which might be better placed here.
Pg 7. I find “explore” a strange choice of wording here “We explore the extent to which people’s ability to retain newly-learned word meanings over time is improved by requiring them to retrieve these word meanings during the intervening period between encoding and a later test.” given this is the pre-registered component. Perhaps it would be helpful to first outline that you’ll explore differences between incidental and intentional learning, before going on to examine the influence of testing on longer-term retention in these training paradigms.
Line 86/7. Further discussion of why it’s of interest to compare incidental and intentional learning would be helpful here. You get to this in the paragraphs starting on line 136 (intro to Exp 1) but I think it would help to lay some of this out at this point. Related to the differences between intentional/incidental instruction, you briefly mention that intentional learning might allow for more strategic processing than incidental learning. Perhaps strategic processing might be particularly important for facilitating access to prior knowledge in the case of synonym learning?
Lines 99-101. “If the presence of a quick, immediate vocabulary test can indeed enhance learning/retention for incidentally learned vocabulary this could potentially provide a simple method for boosting vocabulary gains from story reading” I can see how this could be useful in an educational setting, but not so much from recreational book reading?
When describing the caveats of the Goossens et al (2014) study, it might be useful to point out that this was a study of children and the results might differ for adults (who might be better equipped to learn more successfully from the richer contexts that stories provide, for example).
Line 128 – You say here that Exp 1 provides “a foundation for the subsequent preregistered experiments to explore learning in more detail” – what do you mean by “more detail”. There’s still a bit of a disconnect between Exp 1 and Exp 2/3 at this point for me. At this point in the paper I’m wondering whether your logic might have been that the inclusion of an immediate test might be a confounding factor in previous incidental studies, and encourage participants to deploy strategies after the story reading phase (making the whole thing less incidental)? Setting up your lines of logic in the introduction would be helpful for the reader to understand the connections between the present set of experiments.
Line 148 – what do you mean by an “efficiency advantage” for incidental learning?
In setting up the hypotheses for Exp 1, you’re not clear on whether these predictions refer to both test points. I’m presuming this was on purpose given the more exploratory nature of this experiment, but perhaps it’s worth noting that?
Experimental design: There’s no mention of how you landed at the sample size of 40 for Exp 1? Is this sample size comparable to Hulme et al (2019)? It would be useful to note whether it is equivalently powered at least.
Why did you use a children’s author? Were these stimuli designed for children?
It would be useful to clarify whether an inference was always required to extract the meaning (as in the “foam” example provided, thus enhancing the incidental nature of meaning comprehension.
Were participants permitted to re-read each page as many times as they liked? Also, you say that positioning of the words in the story was naturally distributed, but it would be useful to know how they were distributed over the pages (i.e., is it possible that all occurrences of the word to-be-learned appeared on the same page?).
Although it’s stated that the questions did not tap into the new word meanings, do you think it’s still possible that these questions could have introduced strategies that focussed towards comprehension/understanding of the new word meanings? At least, inserting questions during the comprehension process might have worked to improve comprehension, increasing metacognitive awareness of the comprehension process, so it could arguably enhance the benefits of intentional learning. This is referred to as “guided reading” in the education world. There’s some discussion of it in Blything, Hardie and Cain (2020 - https://ila.onlinelibrary.wiley.com/doi/full/10.1002/rrq.279). Perhaps worthy of a brief sentence in the Discussion as something to note for future experiment design.
As an aside, I would love to know whether participants’ vocabulary knowledge on the Mill Hill VT predicts their ability to learn (and particularly retain) words in the incidental condition more than in the intentional condition!!  We’ve seen that vocabulary predicts children’s and adults’ ability to consolidate new words, but only in some experiments – it seems to be stronger when ppts are learning from richer contexts, which of course makes sense.
I’m not sure it’s described anywhere why you chose to use both the recall and meaning matching tasks, and whether you expect them to behave in the same way.
Line 313-315 – Given you assigned each new word meaning three distinctive semantic features I was surprised to read that you scored the recall responses as 0 or 1. Why not use a 0-3 scale for each item, so that depth of semantic knowledge could be captured? This would seem to me to be a more sensitive measure of semantic learning than a binary score.
Line 462 “performance will be compared to a conventional explicit learning baseline” – this is a bit ambiguous as to whether the intentional training condition is the same as that used in Experiment 1.
As in Exp 1, there’s no justification of the sample size. It would also be useful to clarify why more participants were used in Exp 2 than in Exp 1.
Line 533 – it would be useful to remind us how many participants were therefore included in the analysis.
Line 545. Apologies if I am missing something obvious, but I’m confused about the meaning matching task issue (i.e., why you used a four choice task for the immediate test but an eight choice task for the delayed test). Was this because the immediate test was administered immediately after the story/intentional training, whereas all items were put together at the delayed test? Why was this not a problem for Exp 1?
Validity of the findings: Whilst I agree with your interpretation of the test by condition interaction for meaning matching (Exp 1), it might also be worth considering the idea that memory reactivations during sleep have been to be preferentialised towards initially weak traces and less supportive to memories that already possess strong and enduring representations. One study, for example, showed a memory benefit of sleep for word pairs learned to a criterion of 60% accuracy but not for word pairs learned to a criterion of 90% accuracy (Drosopoulos, Schulze, Fischer & Born, 2007, JEPG). Thus, this could offer an important alternative explanation for your results here.
Line 731 – This could do with some rewording to tone it down slightly given the non-significant effects… “There was non-significantly less forgetting of items trained incidentally through the stories, and the testing effect was also non-significantly larger for incidentally-trained items, which seemed to benefit from the additional learning opportunity afforded by the immediate test.” (e.g., I think it would be less clumsy to talk about trends in the data that are deserving of further investigation). Also, are these non-significant interactions sufficiently powered?
1009 – 1011 – I agree that the reduced forgetting following incidental learning is really interesting and I am quite convinced by it, but I think you need to tone down the claim a little and note the need for replication given the non-significant interaction in Exp 2. Also, on line 1003 it would be useful to clarify whether the trend for reduced forgetting following incidental learning was present regardless of an immediate test, otherwise it could be the immediate test that’s contributing to the resilience from forgetting seen in Exp 1.
1041 – I *think* we are usually careful to say that overnight changes in memory in 0-24hr test studies could be consequence of both consolidation during sleep and repeat testing. There is, of course, evidence for a benefit of sleep beyond repeat testing in studies employing a 12-12 design (e.g., Henderson et al., 2012; James et al., 2020), and in Henderson et al (2013) where we omitted the immediate test for a subset of the participants and found no difference in retention at the 24 hour test (but this was admittedly in children who may show bigger effects of sleep-based consolidation). We’ve also used train-twice test once designs (e.g., Weighall et al., 2017) to reduce the effects of testing. So, whilst, yes, I agree that “some studies considering the impact of factors such as the importance of sleep for consolidation have somewhat neglected this important aspect”, I would strongly emphasize the “somewhat” and that we have also taken significant steps to address the testing effect in this work. We are also actually examining the testing effect in context of consolidation so hopefully more papers on come on this important topic!
Additional comments: This paper is a thoroughly enjoyable read. I can see it being highly cited and of use to a breadth of researchers. Really looking forward to seeing how this work progresses!


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: LEARNING NEW WORD MEANINGS FROM STORY READING: THE BENEFIT OF IMMEDIATE TESTING
Review round: 1
Reviewer: 2

Output:
Basic reporting: The three experiments are clearly reported, and the introduction makes clear how they fill an important knowledge gap in the literature. The authors have made all data and analysis scripts available; I tested a number of the analyses and was able to run the code without problems.

1) My main suggestion here is that the introduction would benefit from slightly more coverage of the possible mechanisms for retrieval practice in supporting new memories. In lines 93-108, retrieval practice is introduced as an opportunity for further learning, which to me implies re-exposure to the items rather than the possible memory mechanisms that might be involved in retention. I was pleased to see the authors cover this literature in further detail in the General Discussion (e.g., Antony et al. fast route to consolidation), but at least a brief mention of this literature in the introduction would be helpful in guiding the reader to think about the benefits of retrieval practice beyond it being an additional form of encoding/training.

2) The results section of Experiment 2 in particular is quite dense, with lots of separate follow-up comparisons for the cued recall task which make it hard to follow the key findings. It would really help to see these different simple effects and pairwise comparisons presented in a table together to clarify what each is contributing to our understanding.

3) Minor comments:
• Line 152-154 – the initial hypotheses were not entirely clear to me here, the two parts of sentence seem to contradict each other. I think the “comparably” is perhaps intended to be relative to the findings of Hulme et al. (2019), but it reads to me as if it should be comparably good with intentional learning.
• Line 462 – introducing a test of participants’ knowledge immediately after exposure? (not test?)
• Figure 4 - It may be an issue with the upload/download, but the shading in the Immediate test condition is not as clearly distinguished as it is for all other conditions/figures.
Experimental design: The research questions addressed in this paper are very clearly defined from the outset, and the authors situate the studies very clearly in the existing literature. The second two experiments were pre-registered and the authors have reported any deviations from original plans in a clear and transparent manner. The materials are available and have direct links in the text which make it very easy to view them alongside reading the manuscript. I also particularly appreciated the clear and detailed section of their modelling approach, and think it would have been possible to reproduce the analyses even without the scripts available.

I note only minor suggestions here that would help to guide the reader through changes across experiments, and a few details that would help in ensuring the methods are replicable:
• The participant section for each experiment would benefit from a sample size justification (even if this was a practical one), particularly as this changes after Experiment 1.
• Similarly, it would help to make explicit why the inclusion/analysis approach changed from Experiment 1 to 2 (i.e., not including participants who did not complete the follow-up session)
• Did the authors check for overlap in items between the trained words and filler vocabulary task?
• What was the overall premise of the study given to participants? The authors mention that they were not told of a memory test, but I am unclear over how they were introduced to the experiment.
• What were the timing restrictions on completion of the Day 2 session? It is mentioned that participants were excluded for completing the task after the deadline (e.g., line 170) and the range of completion times (line 303), but it is not clear what the acceptable time period was.
Validity of the findings: A strength of this paper is that the three experiments incorporate a number of internal replications that support the authors’ conclusions (e.g., benefit of intentional learning over incidental learning; more forgetting following intentional learning). Sometimes the findings are not quite clear cut, but the authors have been cautious in stating where effects were not statistically significant. As already mentioned in previous sections, the raw data and analysis scripts are available and easy to reproduce.

I am left with two questions over the validity of the conclusions from the present data, and a further two theoretical considerations in the General Comments Section.

1)  The authors find a bigger drop in performance for items learned in the intentional condition versus the incidental condition when tested the next day. This is an interesting result that is not necessarily intuitive, and the authors suggest some possible explanations regarding the richer semantic context (lines 444, 1007). However, it could also be the case that this difference is a function of initial learning level: initial performance was very high in the intentional learning condition and had much further to fall.

2)  To what extent are the authors confident that participants did not treat the incidental task as an intentional learning task, despite no instruction to do so? Adult participants in psychology experiments are often (irritatingly!) aware of task demands and what might be expected of them, and presenting known words with entirely different meanings is perhaps still very salient to a participant who is second-guessing the tasks ahead. Speaking to this, I was surprised that there was no effect of condition order, but this could suggest that participants were suspicious of the learning task in both conditions. Did the authors collect any participant feedback data that might speak to this concern?
Additional comments: I thoroughly enjoyed reviewing this manuscript: the experiments are well-designed, tackle an important gap in the literature, and I appreciated the authors’ transparency in using pre-registration and sharing all materials, data, and analysis scripts.

I have two further theoretical questions that the authors might consider addressing in their discussion:

1) To what extent could the current findings of a testing benefit be a bias rather than a concrete benefit for the items tested (i.e., also driven by the cost of *not* testing half the items)? If memory capacity is limited, then there may not be any greater improvement seen for providing an immediate test for all items. By testing each participant on only half of the items in the immediate test, those items could be more salient in memory and out-compete items not tested and regarded as less important (e.g., van Dongen et al., 2012). In contrast, a between-subjects design could show that the practice of testing actually enhances performance relative to not testing - rather than just biasing which items are remembered. Confirming that this is the case would be an important pre-requisite to determining whether test-enhanced learning provides an absolute benefit in an applied setting. I am not suggesting another experiment here, but wonder whether this is something the authors have considered or whether anything in their data/literature review speak to this question?

2) The authors make an interesting and important point regarding the use of repeat testing in studies of offline consolidation (lines 1037-1053) and, as they note in the previous paragraph, it is not clear the extent to which the testing benefit observed in these experiments relates to consolidation processes. In this regard, it would be more balanced to also note that there are many studies that have observed an offline consolidation benefit above and beyond repeat testing (e.g., those that use an AM-PM design to compare 12-hour periods associated with wake and sleep; and those that also include a further 24-hour test). The present study is valuable in highlighting the importance of considering *both* aspects together, especially following incidental learning contexts which I think has been somewhat neglected, but does not exclude a role for sleep-based mechanisms in previous (or the present) experiments.


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: LEARNING NEW WORD MEANINGS FROM STORY READING: THE BENEFIT OF IMMEDIATE TESTING
Review round: 1
Reviewer: 3

Output:
Basic reporting: The reporting is clear, unambiguous and complete.

There are two aspects that I think need to be worked out more clearly in the theoretical introduction to provide context.

One is that the distinction between incidental and intentional learning seems to be mixed with the distinction of contextualized and non-contextualized learning at times. Perhaps incidental learning is usually contextualized, but intentional learning can also involve context or inferences of word meaning from context (as a case in point, the authors refer to a publication in which we showed benefits of retrieval practice when participants intentionally practiced vocabulary in a sentence context).

Second, and perhaps related, I miss an argument why the testing effect might be different depending on the type of encoding that preceded retrieval practice. The authors give a relatively broad explanation, suggesting that incidental and intentional encoding might trigger different kinds of information processing (one involving more semantic elaboration, one leading to more attention to the word form, ll. 136ff) and that “it is unclear whether the testing effect would provide a similar benefit for vocabulary learned under incidental learning conditions” (ll. 95f). Yet, it remains open how or why these differences in encoding would moderate the testing effect. I think by providing information on relevant cognitive mechanisms of retrieval practice, this question could be introduced in a more informative and more convincing way. Two suggestions that might help build an argument why incidental versus intentional encoding could influence subsequent retrieval practice differently: (1) there is a debate in the literature on retrieval practice regarding the importance of semantic elaboration and/or reinstatement of the encoding context during retrieval (most cited publications are from the labs of Carpenter or Karpicke, for example). You could possibly link these to your argument that the incidental learning condition may have involved more semantic elaboration or a richer encoding context. (2) the meta-analysis by Rowland (2014) and other publications suggest that benefits of retrieval practice are larger when retrieval success during practice is high, and retrieval success may depend on (the success of) prior encoding. Thus, if intentional encoding is more effective than incidental encoding, that could cause the testing effect to be larger after intentional encoding.

Textual suggestions
l. 151: Quote: “Based on the previous research, we predicted that learning of new word meanings would be better for intentional learning conditions, although we expected comparably good vocabulary learning for the incidental learning conditions in line with the findings of Hulme et al. (2019).”
I may misread the sentence, but the two subclauses seem to contradict one another.

ll. 935. “Although accuracy…” maybe double-check this sentence. It refers to an interaction effect but (seems to) describes a main effect.
l.374, should “although” be “however”?
l.461 remove “a”
l.743 “test” should be “testing”
l. 907 “is” should be “it”
l. 1044 Check use of “Although”
Experimental design: This review follows the structure of the three-experiment paper and not all comments clearly fit into one of the three sections, therefore, the majority is posted here and the comments on conclusions and wider interpretation of findings in the general discussion are included under "Validity of findings".

General comment: Enough information is provided on the methodology of the experiments, which appears sound. The statistical models are described clearly and systematically so that, in spite of the large number of models, it is easy to follow how different effects were tested and how models were selected, including the simple contrast analyses done to follow up on interaction effects. Authors also clearly distinguished between preregistered analyses and analyses that deviated from the preregistered analysis plan.

Experiment 1
Results
-	Figure 1: If I understand correctly, the mean percentage accuracy was calculated per participant across a number of trials. Could you include information about the distribution of values of participants in the plot, for example, by showing a violin plot instead of a bargraph, or by including the raw data observations (for example, using the jitter function in R)? I know that this is a remark that is easily written in a review yet costs the authors plenty of time but if I understand correctly what is depicted, then this adjustment would make the plots more informative.
-	There is an order by condition interaction for the data of Day 1 (ll. 374ff). The authors write “This result was most likely driven by participants becoming somewhat fatigued by the second task and this fatigue effect having greater impact on the story reading task, which took longer.”. I don’t find this a convincing explanation because performance was actually (numerically) higher for the intentional condition when that condition was the second task rather than the first task; thus, there seems to be no general fatigue effect, and in particular no such effect after the longer story reading task.

Experiment 2
Methods
-	It speaks for this study that the authors recruited sufficient participants to work with an unannounced second test. Based on the participant section, I thought that the data of 99 participants were analysed but after reading the procedure (l.533), I got doubts whether you analysed data of 99-36=63 participants or 99 participants. Perhaps these sections can be edited to avoid confusion.

Experiment 3
Introduction
A previous study that contrasted cued recall and multiple choice practice tests in vocabulary learning, including interactions of practice test and posttest format, is Nakata 2016 (doi: 10.1515/iral-2015-0022). I think this should be referenced here or in the general discussion.
Validity of the findings: General Discussion
While reading, I found the three experiments only loosely connected (proceeding from Experiment 1 on incidental versus intentional learning to Experiment 2 on testing effects to Experiment 3 on test format effects). The general discussion brought Experiment 1 and 2 together a bit more, but there is no explicit discussion of Experiment 3. Why is that?

I miss a discussion of the specific characteristics of the stories used in this experiment and how these may have influenced how effectively participants learned the new word meanings in the story condition (e.g., that/whether the target words were central to the story and repeated multiple times, whether the stories were interesting/appealing for the participant audience). In turn, (how) does this limit the generalizability of findings?

Speculation: Regarding the interpretation of enhanced forgetting over time in the intentional condition, an alternative interpretation that could be valuable to add is that the intentional condition brought some weakly encoded items just above the recall threshold for the immediate test, while the incidental condition did not (thus resulting in better immediate test results). If these weaker items fell below the recall threshold of the delayed posttest and thus delayed posttest scores were lower than immediate scores for the intentional encoding condition, is it then fair to say that items were more susceptible to forgetting in the intentional condition compared to the incidental condition? (for a discussion of how distributions of item strengths could cause patterns of posttest findings, in the retrieval practice literature, see Kornell et al, 2011, doi:10.1016/j.jml.2011.04.002 ).

Minor: The explanation of benefits of intentional learning as the intentional task involving some kind of multiple choice retrieval practice (ll.993ff), would be interesting to connect to the finding of Experiment 2 that adding the immediate test had a stronger benefit (testing effect) on incidental learning than intentional learning (possibly because intentional learning already involved a limited testing effect?).
Additional comments: I appreciated the clear writing of this paper and the logical structure, and I applaud the authors’ choice for preregistration and sharing their data and materials online in a format that can easily be reviewed. Before receiving this review invitation I had actually seen the materials of the project online, so they do find their way into the research community and I hope that the open access will lead to their use in future research. The stories are well-written and an interesting tool to study incidental word learning.

I recommend a revision before publication of the paper and I consider most of my suggestions minor revisions.


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: FORMALIZED CLASSIFICATION OF EPHEMERAL WETLAND VEGETATION (ISOËTO-NANOJUNCETEA CLASS) IN POLAND (CENTRAL EUROPE)
Review round: 1
Reviewer: 1

Output:
Basic reporting: The English text of the final version should be checked carefully. A number of misspellings are annotated in the attached pdf-version

all other items o.k.
Experimental design: The authors analysed all the relevés recording ephemeral dwarf vegetation in Poland, which can be classified into the Isoeto-Nanojuncetea class. Releves have been selected from the Polish Vegetation data base on the basis of pre-selected 43 diagnostic species. The paper gives an excellent overview over this vegetation type and habitat, sheltering a high number of vulnerable plant species.

Aims of the study are clearly formulated, methods explained in detail. Statistical treatment (a.o. with Cocktail method) is adequate. The accepted syntaxa at the levels of alliance and association result from a logic and objective classification process using sociological species groups. The data are documented in a synoptic constancy table Tab S2, distribution in Poland is given in a series of maps.
Validity of the findings: no comments, but see general comments for the authors
Additional comments: There are two major points, authors should take into consideration for the revision:
1. I would hesitate to describe the stands with Coleanthus subtilis as a new association (lines 615-619) and would prefer to classify it as a rankless community or dominance community. It is of course a clearly separated cluster in your data set, but this new association is only of local relevance in a very small part of Poland. C. s. is scattered locally over Europa, Asia and North America, and can be an ephemeral, locally dominant species associated with different species and occur under a variety of environmental conditions (see Richert et al. 2016). It occurs in various Eleocharition and Nanocyperion communities. When describing it formally as ass. nova, you should compare it with other syntaxa, just to mention in Eleocharito-Caricetum bohemicae in Austria (Koch et al. 2005, Bernhardt et al. 2005), Coleantho-Spergularietum echinetosum Hejny 1978 in CZ, Chenopodio rubri-Coleanthetum subtilis Le Bail et al. 2013 in Foucault 2013 in France, Rorippo-Limoselletum coleanthetosum Taran 2005 in Western Siberia, with Limosella aquatica in Kanada (Catling 2009)
2. You should make clearer in chapters results and discussion, that the differentiating and explaining environmental factors (see ordination diagrams) are concluded from the Ellenberg Indicator Values of the considered species and are not based upon observations or measurements on the plots. And you should discuss, that other ecological factors (not available for the plots) might be more relevant (and less “proxy factors”) as selective filters for the species assemblages (just to mention a few: hydroperiod, duration and depth of submergence, surface temperature during emergence and germination of the annuals etc).

Recommendation for authors and editors: To my opinion, it would be better for the reader to shift the constancy table Tab S2 from the electronic supplement into the printed version

minor comments
Line 127: The number of finally used relevés (816) should be mentioned in the abstract
Lines 180-182 belong to methods
Lines 180-182 refer to Tab 1 in text here
please give the arguments why you propose a new name for Digitario-Illecebretum and refer to the new version of ICPN (Theurillat et al. 2020)


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: FORMALIZED CLASSIFICATION OF EPHEMERAL WETLAND VEGETATION (ISOËTO-NANOJUNCETEA CLASS) IN POLAND (CENTRAL EUROPE)
Review round: 1
Reviewer: 2

Output:
Basic reporting: The revised manuscript “Formalized classification of ephemeral wetland vegetation
(Isoëto-Nanojuncetea class) in Poland (Central Europe) ” is generally well-written, clear, and in my opinion definitely worth to be published. Figures and Tables are necessary and well designed.
This study, although it has rather the nature of checklist, presents interesting data on vegetation classification of rare, protected and definitely under examined plant vegetation from Isoëto-Nanojuncetea class. The authors analysed 1340 relevés (of almost 70000!) that were classified to the I-N class, as well as described and discussed 13 associations (among them Coleanthetum subtilis as a new syntaxon). Besides, they presented the maps of distributions of the associations in Poland and statistical-ecological analyses of the analysed plant communities. Of all the reasons, I find this manuscript very interesting and useful. It share the light on the syntaxonomy of this unique vegetation in Poland (central Europe) and the results will be adopted and used in conservation strategies both in Poland and European Union, having in mind that the habitats is protected under the Natura 2000 programme.
Experimental design: Research questions were well defined and the ms fills the knowledge gap in current and proper taxonomy of this plant vegetation.
Validity of the findings: no comments
Additional comments: The manuscript, in some parts needs supplementation

In my opinion, cluster analyses of the analysed syntaxon should be presented, to show the readers general ordination of this ephemeral wetland vegetation in Poland (in addition to analyses of particular orders).

Particular remarks
Abstract:
Reliable and formalized classification of the class Isoëto-Nanojuncetea has not been
performed in Poland. – I think that it is NOT true! (see works of A. Popiela in References, as well as Kornaś (1960), Wójcik (1968) and Zając (1988))  Please remove or rework this phrase.

line 56 - defined. examples are vegetation of the class Isoëto-Nanojuncetea – remove dot.
line 66 – I think that the most recent literature date on classification of Isoëto-Nanojuncetea should be added.
line 76 - Plant communities of Isoëto-Nanojuncetea class in Polish territory reaches – replace with Plant communities of Isoëto-Nanojuncetea class in Poland reaches
line 86 – add Bruelheide, H., et al 2019. sPlot – a new tool for global vegetation analyses. – Journal of Vegetation Science. 30(2): 161-186. https://doi.org/10.1111/jvs.12710
In the Introduction the authors should mentioned on conservation and the importance of this vegetation in European Union. The threats of particular communities should be also presented/discussed in the MS either in description of particular syntaxa or in the Discussion
Results should starts from the general ordination analysis of the studied vegetation type/analysed vegetation plots. In the description of particular syntaxa, a short information on previous studies of these vegetation will be welcome. Now it looks like all of these associations are studied here for the first time.
line 194 In Poland, the Eleocharition ovatae alliance includes three plant communities – replace included with comprises
line 187 Differential – it will be easier to readers if the taxa in Differential, Constant and Dominant subsubchapters will be presented in alphabetic order
line 188 Constant species: Cyperus fuscus, - The nomenclature of species in the MS should be checked, here is Cyperus and afterwards Pycreus – so please be consequent. Moreover you already stated that you used Euro-Med-Plant+ nomenclature for species names.
line 228 Cyperetum micheliani Horvatić 1931 – I am wonder if this name is OK? I think that the authors should explain why they have no Cyperus micheliana in diagnostic species in Notes. Is this name Cyperetum micheliani shouldn’t be replaced by more actual?
line 253-254 -  Coleanthetum subtilis ass. nova hoc loco; Lectotypus hoc loco designatus: Dajdok et al. (2017) – I do not understand it. This plant ass has been described by the authors in this MS ass. nova hoc loco! – so why you decided to cite lectotype instead of holotype? Or maybe you are going to describe it after Dajdok??? as Coleanthetum subtilis Dajdok ex Kącki et al. ??? I am not familiar with such nomenclature, but it should be explained in the Notes (after Habitat requirements). In Phytotaxa there is a paper on current position of Coleanthus (Nosov et al.) – maybe it also will be useful.
317 Cyperetum flavescentis Koch 1926 - Diagnostic species: Pycreus flavescens - - I think that in this case the name of association could be replaced with Pycreetum flavescentis??
363 Eleocharito-Schoenoplectetum supini Soo & Ubrizsy 1948 nomina inversa prop. – please explain in Notes how it was earlier, and why this nomina has been proposed. By the way souldn’t be now ‘Eleocharito-Schoenoplectelietum supini??’
429 Distribution: Stellario-Isolepidetum – please do not use abbreviations here and leater.
441 Centunculo minimi-Anthoceretum punctati Koch ex Libbert 1932 – I am wonder if it shouldn’t be now Anthocero punctati-Anagaletum minimi Koch ex Libbert 1932 or Anagalo minimi-Anthoceretum punctati Koch ex Libbert 1932??
 495 Digitario ischaemi-Illecebretum verticillati Diemont & al. 1940 nom. mut. propos. – please explain in Notes why
582 European range (Popiela 2005). Moreover, a large part of - -  Moreover, a majority of


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: FORMALIZED CLASSIFICATION OF EPHEMERAL WETLAND VEGETATION (ISOËTO-NANOJUNCETEA CLASS) IN POLAND (CENTRAL EUROPE)
Review round: 2
Reviewer: 1

Output:
Basic reporting: see general recommendations
Experimental design: see general recommendations
Validity of the findings: see general recommendations
Additional comments: the revised version can be accepted after some minor linguistic corrections (see annotations in attached pdf)


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

Input:
Title: FORMALIZED CLASSIFICATION OF EPHEMERAL WETLAND VEGETATION (ISOËTO-NANOJUNCETEA CLASS) IN POLAND (CENTRAL EUROPE)
Review round: 2
Reviewer: 2

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: I find the ms sufficiently improved and it can be published in its current form. I have no further comments to the manuscript. The authors should however consider if the community of Coleanthus subtilis needs to be lectotypified (?).


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

Input:
Title: THE EFFECT OF DIFFERENT SOWING DATES ON DRY MATTER AND NITROGEN DYNAMICS FOR WINTER WHEAT: AN EXPERIMENTAL SIMULATION STUDY
Review round: 1
Reviewer: 1

Output:
Basic reporting: Fig.1 , X axis indication are missed, please report date in term of month, if the months are considered. /
L166-171. You describe the plant samples and the N determination. It is clear that you sampling the different plant part, leaves, stem etc, what is not clear and not specified if you determine the N on the single parte , i.e. leaves, or if you , after dry mix the different part to get a single samples. The procedure is not clear, even if later you report that the N accumulation is the sum of the different organ. Please rewrite this paragraph .
Check caption table 5-7

In several part there are spelling error, e.g eigen value =eigenvalue
Experimental design: no comment
Validity of the findings: no comment
Additional comments: General comment
The manuscript is interesting but some section are not clear in English. Therefore, I suggest a English revision and an improvement in clarity in material and methods. This minor revision will make the manuscript better in quality and able to be publish.


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

Input:
Title: THE EFFECT OF DIFFERENT SOWING DATES ON DRY MATTER AND NITROGEN DYNAMICS FOR WINTER WHEAT: AN EXPERIMENTAL SIMULATION STUDY
Review round: 1
Reviewer: 2

Output:
Basic reporting: 1. Manuscript is written in standard English.
2. Literature References are insufficient and some of them are too old (references from 1959 is mentioned).
3. Article structures need to be improved, figure resolution is to be improved.
4. Result and Discussion section is also need to be improved.
Experimental design: Does not meet entirely.... Needs improvement (specifically shared with Author's section).
Validity of the findings: Result and Discussion need to be improved (specifically shared with Author's section).
Additional comments: The research article entitled Different sowing dates effect on dry matter and nitrogen dynamics for winter wheat: An experimental study based on simulation is a quite well documented approach by Liu et. al., in terms of assessing the effects of different sowing dates on different agronomic parameters of winter wheat and checking relevance of a logistics model in quantifying Dry Matter accumulation (DMA) and Nitrogen Accumulation (NA) in winter wheat. As the manuscript was observed further, some noticeable rearrangement issues of this article are needed to be rectified. These are as follows-
1.	In the Abstract section, please mention the novelty and objectives of this research work in the ‘Background’ part and mention the variables of the experiments for each objectives within the ‘Methods’ section. Also mention the novelty in clear cut sentence within the “Introduction” portion as well.
2.	Please provide the full form of all of the abbreviations during their first mention, and then apply abbreviations within the text. Please also write ‘Nitrogen Accumulation’ in place of ‘N Accumulation’.
3.	Please provide the initial physicochemical characteristics of the soil in a separate table.
4.	Please provide the bibliographic reference to the fertilizer application practice followed. Please also explain in brief why an additional 90 kg ha-1 of N was applied at the beginning of the jointing growing stage?
5.	Please provide weather and climatic information (max-min temperatures, RH and rainfall pattern and daily sunshine hour) with proper bibliographic refetence, if not recorded by researcher.
6.	Please also prove the fertilizer application history/pattern of that area (include previous crops also).
7.	Please mention ‘Experimental Design’ properly… whether its CRD/RBD or Split plot… Please mention that.
8.	Please also provide suitable bibliographic reference to the analytical procedures followed in Measurement items and methods.
9.	Please mention the version of M.S. Excel.
10.	Please mention and provide tables for the information mentioned under ‘Weather conditions and crop phenology’ and ‘Morphological traits’.
11.	Please increase the resolution of the figures and make them bigger in size so that each detail is easily observable.
12.	It is highly appreciable if the authors could improve the discussion section. The entire Discussions section is highly inconsistent in relating the results with the objectives to be fulfilled. Please take care of the following points within the Discussion segment-
a.	Please address the findings of the Result section in relation with the fulfillment of the objectives.
b.	Please add more relevant references of previous works and relate the current research.
c.	At the end of the discussion, mention in clear cut sentences, how far the objectives are fulfilled and also mention of there is any future scope of research.
d.	Write the conclusion mentioning the main points from the reformed Discussion segment.
13.	It is really appreciable to observe the low plagiarism of the manuscript (9%) as detected by Plagiarism Checker X (report attached).


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

Input:
Title: POTENTIAL OXYGEN CONSUMPTION AND COMMUNITY COMPOSITION OF SEDIMENT BACTERIA IN A SEASONALLY HYPOXIC ENCLOSED BAY
Review round: 1
Reviewer: 1

Output:
Basic reporting: Yes, English meaning and grammar is clear. One general comment is that there are a lot of abbreviations and keeping track of them mentally is challenging and detracts from the readers capacity to interpret the research. If you are not restricted by word count I would suggest limiting their use by using whole words.

Literature of the use of INTR is out of the scope of my expertise.
Literature on sediments and microbial communities is relevant.

Article is structured into Introduction, methods and combined results/discussion

Figures and tables support primary results. Figure 5 labelling – hexagon would be more appropriately named star. Sequencing data is available
-
Bacterial names need to be italicized – see legend Fig 6 and throughout text

More specific comments:
The strength in this study was in the reporting of the chemical data. Associating changes in the composition of the microbial community to the biological consumption of oxygen and the chemical consumption could be improved. My interpretation is that the biological (BOC) is a measure of direct oxygen consumption and that chemical (COC) is a measure of indirect biological consumption mixed with other reductants present within the sediments. With this in mind I find Figure 2 interesting as it implies that the majority of oxygen consumed by the sediment is not directly via respiration (microbial or microphytobenthos). The primary process reported here (but not discussed at length) appears to be via indirect biological processes that increase the proportion of reduced compounds (pCOBINT), which then consume available oxygen. Although difficult to achieve I would have liked the authors to discuss how these data, especially changes in the composition of the microbial community, could be contributing to this. For example, does increased pBOCINT co-occur with a decreased relative abundances of known anaerobes (who could be producing the reduced products)? or in identifying and temporally tracking the strict aerobes (which might be a simpler way to achieve the same thing)?

Line 393-396. Do you have any taxonomic support within your dataset from 2012 for this statement “As the highest pBOCINT in 2012 and 2013 (2.82 µmol INTF g−1 day−1 in 2012 and 4.16 µmol INTF g−1 day−1 in 2013) coincided with the timing of the highest bacterial count in each year (1.7 × 1011 cells/g sediment in 2012 and 3.2 × 1010 cells/g sediment in 2013), it was likely that the increases in facultative anaerobes or aerobes contributed to the surge of the pBOCINT.” One way to analyze this would be to apply a time-series model.  This could be achieve using a log-ratio transformation or a random forest approach?

Line 458 – I agree that making these linkages is important for the interpretation of this study and doing it more would increase the conclusions and relevance to the stated question. However, changes in relative abundances between treatments can not be compared directly without controlling for change in the relative abundances of taxa around them. Although with their own biases, tools exist to achieve this such as deseq and ancom. See: “It was interesting to note that the increase in the relative abundance of Arcobacter ASVs coincided with the highest pBOC (Fig. 2).”

Figure 6 – Instead of labelling by month sampled it would be interesting to label these data with the oxygen availability in situ. As the impact of oxygenation after 24h will change based on the pre-existing oxygen conditions of the sediment.

Line 62 – define NH4+, Fe2+, and HS-,
Line 127-129 – is this required? (This research was conducted by Fumiaki Mori in partial fulfillment of the requirements for a Ph.D. from Nagasaki University (Mori, 2018))
Line 385 – which part of fig 2 are you referring to here?
Line 396 – add table 1 reference to bacterial cell counts
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: POTENTIAL OXYGEN CONSUMPTION AND COMMUNITY COMPOSITION OF SEDIMENT BACTERIA IN A SEASONALLY HYPOXIC ENCLOSED BAY
Review round: 1
Reviewer: 2

Output:
Basic reporting: The manuscript is clear and professional. Some minor observations:

Line 29, including the frequency of the monitoring.

Line 60: grammatical error, change "in" to "of"

Line 151: delete “of”

Line 357: “BIR” is not defined in the whole document.
Experimental design: In the equation in line 183, it is unclear if the incubation time changed between the experiments.

Line 190, ¿What kind of bottles? ¿ODB, PC?

The GAM that was adjusted is not shown. I suggest that the authors show the GAM equation just as they did with the GLM (Eq. 2).

Line 238: What is the proportion of isopropanol and methanol?

Line 240: Delete: “(See Comparison between….)”
Validity of the findings: Line 326-327: Include the reference for continuous cultures of natural communities of marine prokaryotes: Villegas-Mendoza et al. 2019. "The Chemical Transformation of the Cellular Toxin INT (2- (4-Iodophenyl) -3- (4-Nitrophenyl) -5- (Phenyl) Tetrazolium Chloride) as an Indicator of Prior Respiratory Activity in Aquatic Bacteria" ·.

The authors reasonably argue in lines 342-350 how the incubation of sediments with INT circumvent the limitations (toxicity, INT recovery) reported for the quantification of INTF in planktonic samples.

Line 372: Why did the effect of formalin only occur in 5 of the 20 samples?
Additional comments: The manuscript includes relevant information that contributes to the quantification of respiratory processes in sediments. The relationship of temperature and preservation of organic matter during anoxic conditions seems very interesting (Fig. 3). Although it is mentioned that the temperature is a determining factor of the oxygen consumption of the sediments. It would be enriching for discussion by including a more detailed explanation of the effect of temperature and substrate concentration as a driver of metabolic parameters in the identified microbial community.


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

Input:
Title: POTENTIAL OXYGEN CONSUMPTION AND COMMUNITY COMPOSITION OF SEDIMENT BACTERIA IN A SEASONALLY HYPOXIC ENCLOSED BAY
Review round: 2
Reviewer: 1

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


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

Input:
Title: POTENTIAL OXYGEN CONSUMPTION AND COMMUNITY COMPOSITION OF SEDIMENT BACTERIA IN A SEASONALLY HYPOXIC ENCLOSED BAY
Review round: 2
Reviewer: 2

Output:
Basic reporting: I appreciate the author’s effort in providing answers to my assessment.
Experimental design: The experimental design is robust and detailed. The author has considered the previous comments. The methods provide specific information for their replication.
Validity of the findings: The authors added the information requested in the previous review. The discussion and conclusions are clear and concise and correspond to the information provided in the results.
Additional comments: I appreciate the author’s effort in providing answers to all my questions.


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: ONLINE SEARCH TRENDS AND WORD-RELATED EMOTIONAL RESPONSE DURING COVID-19 LOCKDOWN IN ITALY: A CROSS-SECTIONAL ONLINE STUDY
Review round: 1
Reviewer: 1

Output:
Basic reporting: The authors produced an interesting analysis on the language adopted by Italian people during their first lockdown through Google searches in combination with self-reported metrics of emotional well-being. Through a detailed and careful statistical analysis, the authors found that during the lockdown online searches on Google from Italian people shifted to higher frequency, more concrete, more positive and less arousing concepts. During confinement, individuals reported emotional states indicating a general loss of sense of agency and potential fearful states. Interestingly fear modulated different emotional dynamics in individuals. Those less subject to fear perceived concepts with less arousal, with additional losses of valence and dominance only in relation to COVID-related terms. Those people more prone to fearful states, ended up perceiving with less valence and dominance not only COVID-related but also COVID-unrelated concepts, a sign that the Italian lockdown provided enhanced emotional damage to the wellbeing of those people more sensitive to fear or other negative emotional states.

The manuscript is very well written, concise and clear. The topic is definitely of crucial relevance, given the need and the difficulty to quantify the emotional repercussions of lockdowns over individuals’ wellbeing. The authors have to be praised for their multidisciplinary approach, declining statistical techniques frequently used in psycholinguistics to a more social- and complexity-related scenario like the global pandemic of COVID-19. For these reasons I recommend the publication of this manuscript with minor revisions.
Experimental design: Lines 286 – 311: The report of the statistical testing is correct but it might confuse readers. Would it be possible to translate the information in text into a concise table to include in the main text? At the end of this section it would be useful to add a paragraph summarising the above tests in plain language, in order to better highlight the dynamics of affective ratings under the lockdown.
Validity of the findings: The Discussion could include a few more links and ties of the authors’ interesting results and those of the ever-increasing literature on COVID-19 and language analyses. I reported 3 main relationships that might be of relevance for the authors’ work:
- Dyer and Kolic (APN, 2020) found that online media indicated psychophysical numbing in people subject to restrictive health measures across 12 countries. Social discourse results being decreasingly emotional and increasingly analytic when discussing COVID-related jargon. This change detected in the language used by individuals could be interestingly connected with the emotional alterations identified by the authors, particularly in those individuals not being strongly exposed to maladaptation.
- Mazzuca et al. (PsyArxiv/4ndb8, 2021) found that the first Italian lockdown altered drastically the conceptual representation of concepts related to COVID-19, boosting the occurrence of conceptual associations from the illness domain like “flu-fear” or “disease-mask”. Could these shifts in the mental representation of the concepts be reconciled with the finding reported by the authors in this analysis? This connection would be interesting also in view of the authors’ analysis on how different individuals responded with different changes in their perceived emotional features of concepts.
- Stella (First Monday, 2020) investigated the language produced by Italians at the end of the first lockdown, in view of national re-opening. The analysis found that Italians tended to share more messages including a higher content in fear. The act of resharing is not always an endorsement but it is strongly triggered by a user sharing a similar view or an interest in the reshared content. In this way, the authors’ finding of improved feelings of fear and reduced sense of agency might be the source for the behavioural pattern found online in this paper although more research would be necessary in that direction.
Figure 1 – It seems to me like “hotel” was chosen as an example word for statistical reasons. However, the decrease of search occurrences for “hotel” is key to a more limited mobility being suddenly imposed to Italians. It might be worth mentioning this in the main text, especially since “hotel” are now being increasingly associated with COVID-19 (COVID-19 hotels are structures where quarantine or asymptomatic individuals can spend some time).
Additional comments: No additional comments