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 IMPORTANCE OF OPEN SCIENCE FOR BIOLOGICAL ASSESSMENT OF AQUATIC ENVIRONMENTS
Review round: 1
Reviewer: 3

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: This is an important and innovative paper that highlight the issue of the accessibility of scientific results and data to facilitate the bioassessment of environmental impacts. The paper is very timely as the open science is becoming a standard for most of biological studies and it’s important that this practice is also adopted by ecological studies of practical importance for environmental managers.

The review is very well written, and I fully recommend its publication. I would suggest however few points to be considered in the revision of this paper.

The first point concerns the new ecogenomic tools that are currently transforming the field of biomonitoring and bioassessment. I was quite surprised to find no reference to these tools despite the fact that their impact on so-called “democratization” of bioassessment practices is well documented (e.g. Deiner et al. 2017, Pawlowski et al. 2018, Hering et al. 2019). In particular, it is worth to be mentioned that the access to genomic or metagenomic data can be much easier and more open compared to the classical morphotaxonomic data, which are more depending on personal expertise and more difficult to compare with others. The authors could address this point when they are discussing the issue of species identification (subchapter Curating bioassessment data).

The second point, somehow related to the first, concerns a certain conservationism of bioassessment practitioners regarding scientific progress. As discussed by Guareschi et al. 2019, this is not only about new tools but also about ecological changes, such as the presence of alien species, or changes in taxonomic identification. Of course, open science can help pointing out these changes and interpreting them correctly. However, even the most accessible research will do nothing if the environmental managers are not particularly interested in getting updated about the new research advances and implementing them in their practice.

My final critical remark concerns the scope of the review and its emphasis on open source software applications. The authors provide a very exhaustive description of using R for bioassessment, but they go into a lot of details that somehow contrast with a general character of the review. They assume most of environmental managers know how to use R, which I doubt is true. In fact, the authors seem to agree with me, when they point out that the education and training are the main limitations of the access to the open science tools and data. It’s probably the main message of this review paper and I would rather focus on it than on describing the various applications of tools that most probably remain inaccessible for most of practitioners.

References:
Hering et al. 2019
https://www.sciencedirect.com/science/article/pii/S0043135418301830?via%3Dihub
Guareschi et al. 2019
https://www.sciencedirect.com/science/article/pii/S0048969719300087?via%3Dihub
Deiner et al. 2017
https://onlinelibrary.wiley.com/doi/full/10.1111/mec.14350


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 IMPORTANCE OF OPEN SCIENCE FOR BIOLOGICAL ASSESSMENT OF AQUATIC ENVIRONMENTS
Review round: 2
Reviewer: 1

Output:
Basic reporting: The authors have addressed the concerns raised well. I think the article is a timely, well written, and informative addition to the literature that I would like to recommend this for publication.
I have a few mirror comments with regards some of the new sections that the authors may wish to consider, if they are given the opportunity, but I do not think these changes are completely necessary.
In the new section describing pre-registration (from line 264), it might be worth mentioning that some journals offer grantee of publication (if protocols are stuck to) irrespective of null findings. This is what prevents publication bias, not pre-registration per se. Most journals differentiate between pre-registration (which guards against other questionable research practices such as p-hacking and HARKing) and Registered Reports which are a type of pre-registration but with the additional guarantee and peer reviewer from a journal prior to data collection. If the authors wishes to include this differentiation I think it might help the readers understanding. However, I appreciate that these terms are often used interchangeably. I also appreciate that this does not site perfectly with the scheme set out in figure 1, but as Registered Reports are becoming more broadly accepted, with over 240 participating journals, including many specialising in ecology (see https://www.cos.io/our-services/registered-reports?_ga=2.14444030.83877480.1591349264-1270378436.1498895408), I would recommend mention of them.
Also, the authors differentiate between exploratory and applied research implying that pre-registration could be useful for declaring research to be exploratory (lines 277-280). I found this confusing, although again this is likely to be subject specific. Commonly, amongst the open science community, the relevant distinction is between exploratory (i.e. not pre-registered, where data is explored) and confirmatory (pre-registered) analyses.
Experimental design: Fine - as before.
Validity of the findings: Fine - as before.
Additional comments: Fine - as before. See basic reporting comment.


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

Input:
Title: THE IMPORTANCE OF OPEN SCIENCE FOR BIOLOGICAL ASSESSMENT OF AQUATIC ENVIRONMENTS
Review round: 2
Reviewer: 2

Output:
Basic reporting: I have carefully read the responses to my comments and those from the other reviewers, as well as the new version of the manuscript and the reviewers have addressed satisfactorily them. The new version of the manuscript has improved and now can be published.
Experimental design: the comments have been addressed adequately
Validity of the findings: the comments have been addressed adequately
Additional comments: No additional comments


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

Input:
Title: APPLICATION OF MATERIAL FROM USED CAR TYRES IN GEOTECHNICS—AN ENVIRONMENTAL IMPACT ANALYSIS
Review round: 1
Reviewer: 1

Output:
Basic reporting: 1.	Manuscript presents results about important issues of environmental impact of recycled tyres, in particular the use of tyre bales.
2.	Paper was written well with good English. Design of the paper is easy to follow.
3.	Figures are in good quality, however it is hard to see what figure 4b) presents.
4.	Tables are clear and complete.
5.	References should be improved with the recent one.
Experimental design: 1. This research is consistent with the aims and scope of the journal. It contains environmental issues connected with tyre recycling.
2. The research goal was clearly stated at the end of the introduction.
3. Authors presented wide literature overview, however the part connected with leaching of different substances (lines 95-147) was written on the basis of old references. It should be improved with recent achievements in this field.
4. In general introduction is written well, but the research gap filled should be underlined. I recommend to expand the introduction with information about using the tyre bales. Was there any previous publication about use of tyre bales and their environmental impact?
5. Materials and methods section describe experiment in detail, however some explanation and additional comments are needed:
a) What tyres was used in the experiment? Was only one type of tyres used or mixed types? Were they from passengers car or from trucks? Which sizes where they? Depending on the tyre type there will be some differences in the materials used in the design of the tyre, at this can affect the substances leaching and may be important in the context of the results.
b)It will be good to shortly characterize the methods used in analyzes of environmental impact. They were presented in the table 3, but short description of the methods used, in the context of repetition and sample size, will be useful.
6. The results section presents interesting results describing environmental impact of tyres bales in the context of leaching. Authors clearly present the complete results. The are, however, some issues need to be improved:
a) the discussion and comparison of the results with previous study should be done,
b) it is not clear now if the time of staying bales in the water is important for the leaching phenomenon. Why 120 days of experiment was set?
c) the reasons of the increasing or decreasing levels of substances can be described.
Validity of the findings: a)	All underlying data have been provided; they are robust, statistically sound, & controlled.
b)	Conclusions are well stated, linked to original research question & limited to supporting results, however it can be improved according to my comment 6 from Experimental design.
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: APPLICATION OF MATERIAL FROM USED CAR TYRES IN GEOTECHNICS—AN ENVIRONMENTAL IMPACT ANALYSIS
Review round: 1
Reviewer: 2

Output:
Basic reporting: clear
Experimental design: within Aims and Scope of the journal
Validity of the findings: innovative
Additional comments: In this paper, the environmental impact analysis from used car tyres in geotechnics was conducted. This research is innovative. But some issues should be solved:

Introduction: Authors should remove unnecessary information. For more information regarding generation and utilization of on waste rubber and more application of rubber tire, you should refer and cite the following paper:

Liu, L., Cai, G., Zhang, J., Liu, X., & Liu, K. (2020). Evaluation of engineering properties and environmental effect of recycled waste tire-sand/soil in geotechnical engineering: A compressive review. Renewable and Sustainable Energy Reviews, 126, 109831.

Liu, L., Cai, G., & Liu, X. (2020). Investigation of thermal conductivity and prediction model of recycled tire rubber-sand mixtures as lightweight backfill. Construction and Building Materials, 248, 118657.

Liu, L., Cai, G., & Liu, S. (2018). Compression properties and micro-mechanisms of rubber-sand particle mixtures considering grain breakage. Construction and Building Materials, 187, 1061-1072.

Tyre bales: Description for the tyre bales is not clear. Please add more information.

Results and Discussion: Authors were suggested to add the approximate reason for the results.


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

Input:
Title: APPLICATION OF MATERIAL FROM USED CAR TYRES IN GEOTECHNICS—AN ENVIRONMENTAL IMPACT ANALYSIS
Review round: 1
Reviewer: 3

Output:
Basic reporting: Application of material from used car tyres in geotechnics - environmental impact analysis

the paper met the standards of the journal
even though the writing is ambiguous confusing in some parts
the the results shows interesting alternate material
Experimental design: the experimental method is  adequate
and investigation performed to support the objective
Validity of the findings: the findings are good and recycle the waste tyres is though not novel its impact on environment is being analysed
Additional comments: The purpose of the research carried out and described in this article has been to determine whether tyre bales cause pollution of rainwater, and then groundwater and soil; as well as what level of leaching of undesirable substances

This should be highlighted in the abstract itself

149 paragraph is  repeated
172 Tyre bales are potentially a new building material from the recycling market in transport-  confusing
modify
apart from that ensure that the format of the journal is maintained


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: POTS VS TRAMMEL NETS: A CATCH COMPARISON STUDY IN A MEDITERRANEAN SMALL-SCALE FISHERY
Review round: 1
Reviewer: 1

Output:
Basic reporting: The reporting style is good with clear headings and concise text. The use of tables for data and results avoids an unwieldy results section. There are a couple of niche terms that are used (pots, soak time) that may need to be altered throughout to reach a wider audience. Citations are relevant and sufficient, although clarification is needed one of the comments made in association with the discard ban.
Experimental design: The experimental design is clear, and the aims of the study are clearly highlighted. The requirement for the research is highlighted. There are a differences in sample size and number of hauls at each location, these whilst accounted for in the analysis by standardising the CPUE data, still need explaining for the reasons behind the difference. The description of the gear type used is excellent and can be easily replicated by other researchers. The anlaysis is suitable and in line with other research of a similar field.
Validity of the findings: The validity of the findings and conclusions the authors have made are suitable and also the limitations of the research are clearly defined. The raw data is provided and of sufficient detail to suit the analysis conducted.
Additional comments: I have enjoyed reviewing your research and the relevance of such with the current agenda to reduce discards under the landing’s obligation. At sea sampling designs associated with commercial fisheries can often be difficult to achieve and this manuscript adds to this field well.
A minor point is the use of the word ‘pots’ to describe your alternative method to trammel nets. I recognise pots as a term used for predominantly crustacean fisheries, developed from the more traditional term creel. In this experiment I suggest the term ‘trap’ is used as this is more widely recognised when targeting fish (and cephalopods) as opposed to crustaceans. Fish traps are widely used across diverse fisheries globally, using the term ‘pot’ may restrict the research from reaching a wider audience.
Below are specific comments to the manuscript:
Abstract
Highlight the problem with high discard rates – many discarded species have high discard mortality.
Line 24: Change “more sustainable gears” to “a more sustainable gear type”
You discuss gill nets and trammel nets in the abstract, although they work the same way they are different, it is more suitable to discuss one net type in the abstract to avoid confusion.

Introduction
There is a bit of confusion when describing the gillnets and trammel nets, as I understand it, although both gear types “gill” the fish, a gillnet is a single sheet whereas a trammel net tends to be three sheets – a small inner mesh panel and two larger mesh outer panels. The description in the introduction needs to be clearer in highlighting the differences.
Line 43:  Change “for about” to approximately”
Lines 58 – 60: Is there legislation that sets mesh sizes in these fisheries which may account for high discard rates?
Line 82: “pots have a low energy footprint” this statement is not necessarily true. There has been recent literature highlighting that in pot fisheries the carbon footprint can actually be higher than in mobile fisheries. See:
Abernethy, K.E., Trebilcock, P., Kebede, B., Allison, E.H. and Dulvy, N.K. (2010). Fuelling the decline in UK fishing communities? ICES Journal of Marine Science. 67(5), pp.1076–1085.
Arnason, R. (2007). The economics of rising fuel costs and European fisheries. EuroChoices. 6(1), pp.22–29.
Lines 102 – 106: It took several reads to understand what you are trying to say – this section could do with clarifying.
Material and methods
124 – 126: Do you think the spatial proximity of the GTR’s and fish traps may have affected the results?
Line 134 – 135: Why is the net length at Senigalla a different length – add reason for this.
Line 146: Again highlight why the number of pots differed
Line 147: Define soak time for the nonexpert reader
Line 165: Why was a Kruskal Wallis test chosen – if the data required a non-parametric test or violated the assumptions of a parametric test, this needs stating.
Line 179: What was the protocol used for AIC model selection – lowest value? If so, this needs stating.
Line 180: Was there any correction for soak time?
Results
Line 192: Change “the nets” to “GTRs” – maintain consistency in reporting
Line 198: Change “but especially” to “were more abundant”
Discussion
Line 237: There are many species that have a defined MCRS that are not subjected to the discard ban – Homarus gammarus, Cancer pagurus etc. Needs rephrasing
Line 246: What pots used in the area? By other fishers? Explain
Line 256 add a comma after “study”
Line 257 add a comma after “catch”. What is the value of these numbers? Individuals caught? CPUE? How does this stress a need for bait – clarification needed.
Line 262: Is there high discard mortality to increase the impact on the benthic community?
Line 270: Remove “in” after the word several
Line 271 – 272: This sentence is confusing, reword for clarity
279: Change “bigger” for “larger”

Figure 1: Why is there more points for M. Ravenna than the other sites? Was most of the sampling done at this site and only done once at the others? A smaller scale will highlight the sampling stations better as you have a good inset highlighting the general location.
Figure 2: The description in the methodology section of the trammel net design is excellent and really clear – this figure does not reflect the description. Maybe add a bit more detail to the legend, i.e. what the abbreviations stand for.
Figure 5: It would be helpful if the MCRS for each species could be plotted on the LFD so the reader can see the CPUE of the discards/catch between pot types.
Figure 6: As with figure 5. Does the dotted and dashed line in the shaded areas represent the mean of the data? What does the vertical dotted line at 12 cm denote on the D. annularis plot represent? A label plotted on both plots highlighting on which side of the 0.5 line the data represents Pots and which side represents GTRs.


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: POTS VS TRAMMEL NETS: A CATCH COMPARISON STUDY IN A MEDITERRANEAN SMALL-SCALE FISHERY
Review round: 1
Reviewer: 2

Output:
Basic reporting: I have not offered a detailed response in this section given the issue highlighted in section 3.
Experimental design: I have not offered a detailed response in this section given the issue highlighted in section 3.
Validity of the findings: The authors approach to data transformation is of significant concern and in the reviewers opinion will require comprehensive revision and full reassessment. Unfortunately I believe the current approach has undermined the paper and the fundamental validity of the results reported.

The authors have presented data transformation and standardisation of CPUE for GTR to 12 hrs per 1000m (L158 – L16) and Pots to 24 hrs per 1000m (L161 – L163). Whilst I appreciate the basic narrative and apparent attempt to align with industry working practices, this transformation of data to incomparable temporal scales is statistically unsound and will have fundamentally influenced the proceeding complementary analysis, weighting a significant negative bias against GTR catches in the subsequent comparisons against fish traps. As this data forms the foundation of the manuscript, this issue will have influenced all proceeding data analysis, results and the conclusions presented.

I would strongly recommend the authors redevelop their analysis and compare catches from both gears to a standardised period prior to any resubmission. The scientific question and driver behind this study is about the relative catch efficiency of different gears types, not actual industry working practices, however this could be assessed through a seperate complementary analysis within a revised manuscript.
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: POTS VS TRAMMEL NETS: A CATCH COMPARISON STUDY IN A MEDITERRANEAN SMALL-SCALE FISHERY
Review round: 2
Reviewer: 1

Output:
Basic reporting: Some literature references should be add in the Introduction section.
Experimental design: Standarization of captures should be carefully reviewed.
Validity of the findings: Some results should be added and clarified.
Additional comments: This study contributes to the general interest to the advancement of applied fisheries science. Discard amounts is currently a major concern for the EU Policy and the development of new techniques aimed at reducing discards is necessary.
However, several aspects need be improved in the manuscript. Mainly, Results section should be clarified and some additional results should be added.
Abstract:
Line 30: the two most abundant commercial species.
Introduction:
Line 46-49: Information about how set nets catch fish seems unnecessary.
Line 52: Replace “Amengueal-Ramis et al., 2016” by “Amengual-Ramis et al., 2016”.
Line 57: You could add other studies about discards of trammel nets in Mediterranean sea.
Line 62: Here you could also try to add other references.
Line 68-72: Also the Common Fishery Police in the European Union has focused on the discard reduction. You talk about in the Discussion section but you could add some information here too.
Line 110-112: You should add more details about the aims of the study. For example, to test the effectiveness of the pots reducing discards, …
Materials and Methods:
Line 122-123: The information about the bottoms that appears in the text is not the same as in the Table 1. What is the correct information? Also, you do not need to provide this information in both places (text and table), it is too repetitive.
Line 125: Three different vessels in the three study areas?
Line 126: Why “traditional trammel net” is abbreviated as “GTR”, not as “TTN”?
Table 1: In addition to the mean value (AVG), you should add a standard deviation, standard error or range.
Line 127: I’m not sure the term “set” is clear enough. Do you mean “fishing trips” or “fishing journeys”?
Line 135: Is the diameter of float line 5.5 mm as you say in the text or 8 mm as you say in the Figure 2?
Line 137-138: The sentence “the hanging ratio of...” is repetitive because you repeat all this information in the Figure 2. You could delete it.
Line 139: “and 300 m at Senigallia and”. Delete the second “and”.
Line 141: Is “Trapula” the name of the factory? Clarify this.
Line 154: Each fishing journey, what do you haul? A trammel net, a line of small pots and other line with large pots? Clarify.
Line 157: Change by “invertebrates and fish species with no commercial...”.
Line 163-170: I have some doubts about the standardization. I understand that different gears need different soaking times. However, if the pots are commonly retrieved after 2-3 days, why you standardize in 24 hours? And other reflexion, I wonder what is the reason for leaving pots during 2-3 days, if they obtain an amount of commercial captures similar than the trammel nets just in 24 hours? Does the catch really double in 48 hours? Maybe pots need more than 48 hours to start capturing target species, because the first species captured act as a claim. If that is the case, and captures do not occur linearly, standardization in 24 hours would not be valid. If a wide range of soaking time is available, you can test the trend (amount of captures vs soaking time).
Line 175-185: You could clarify this part. What type of pot? Do you analyze all the pots together? Also, you should define each parameter in the equation.
Supplementary Tables: In line 157, you say that discards are that species with no commercial value and individuals under the MCRS. However, in Supplementary Tables, individuals under the MCRS are included in the commercial catches. You should separate the portion that belongs to discarded fraction.
Also, you could highlight (maybe in bold) in the Supplementary tables, the most important species commented in the text.
Line 202: Change “important” by “abundant”.
Line 207-208: S. umbra at Marina di Ravenna is mentioned twice.
Line 210: You should indicate that the mean biomass value is calculated as CPUEw.
Table 2: In the Table caption, you should indicate that the values presented are the mean value, +/- standard deviation or standard error, and the confidential intervals in brackets.
Also, the total values (shown as a separate table) could be added under the three study sites, as a new file.
Line 211: “did not overlap” is not correct. The CI values of the three gears overlapped.
Line 214: The same analysis made with the CPUEw of the total commercial catch, could be made with the CPUEw of the most important species (almost with cuttlefish).
Line 221: Again, specify CPUEw in the mean biomass values.
Line 228: “The lines of LPs and SPs were mostly above those of GTRs, indicating a greater catch efficiency.” That is not clear in Figure 5. You should apply an statistical analysis with the CPUEw or CPUEi.
Figure 5: In figure caption, specify the name of the sites. Also, in the legend, replace BP with LP.
Line 231: “were equally efficient only for the larger sizes”. I’m not able to see that in the Figure 6. For me these results are a bit confusing. You could explain more these results. You could also add other more simple analysis, for example, an one way analysis of the length between the three gears. You could also indicate the percentage of undersized D. annularis captured by each gear.
Line 234: “GTR catch curves”? Rewrite this sentence. I wonder if you could add a curve for GTR catches or at least represent its CI.
Discussion:
Line 242: “(CFP Regulation (EU), 2013/1380)(EU Regulation 1380, 2013)”. Correct this.
Line 243: Delete one “of”.
Line 245: You could add some information about the consequences of the landing obligations for the fishers. For example, see Maynou et al 2018, Fishers’ perceptions of the European Union discards ban: perspective from south European fisheries.
Line 253: What does “new pots” means? Are pots similar as used in this study? Also, link this sentence to the previous paragraph.
Line 256: “A significant difference between trammel nets and pots was found for annular seabream”. This is not shown in the Results section.


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: POTS VS TRAMMEL NETS: A CATCH COMPARISON STUDY IN A MEDITERRANEAN SMALL-SCALE FISHERY
Review round: 2
Reviewer: 2

Output:
Basic reporting: The paper of Petetta et al. concerns an interesting work aimed at testing the catch efficiency of two type of pots (still not used in Mediterranean) to reduce discards, to be used as an alternative of set nets.
The work is of interest, because it deals with a current topic, e.g. the reducion of discards (see the current management policies on discard ban) and, more in general, with the reduction of the fishing gear impacts and the related socio-economic implications.

This manuscript is a resubmission, following the revision of two referees.
After a careful revision (also of the rebuttal letter), in my opinion the new version submitted by the Authors fully meet the comments made and the explanations are generally satisfactory.

The applied methodology (see below) is correct, the presentation and discussion of the results is clear and unambiguous.

I made a detailed revsion of the text, tables and figures (including the supplementary tables), including several corrections and comments.
I suggest to carefully revise the text accordingly.
Experimental design: The Authors reported that the text was aimed at replacing as much as possible commercial conditions: I agree on this approach but I suggest to explain clearly (at the beginning of Material and Methods) this aspect: e.g. reporting that the characteristics of the gears (length of GRT and GNS), fishing grounds, fisging operations (fishing time), sorTing of the catch, were carried out following the fishermen procedures, without interferences from the scientists on board.
 Of course this approach implied the calculation of different CPUE for the different gears used (I agree that it is not possible to use the same CPUE for gears operating in different ways): I suggest to mention this aspect in the discussion of the results.
Validity of the findings: In my opinion the findings of this study are interesting and can represent an important information for he management of the small scale fisheries and of the coastal fishery resources.
Additional comments: I attach a revised version with  corrections and comments of
- the text (I used the Word revised version - I used the track change)
- the tables and the figures (in the pdf file of the manuscript)
- the two supplemetnary tables (with track 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: POTS VS TRAMMEL NETS: A CATCH COMPARISON STUDY IN A MEDITERRANEAN SMALL-SCALE FISHERY
Review round: 3
Reviewer: 1

Output:
Basic reporting: Some minor modifications are nedeed.
Experimental design: No comment.
Validity of the findings: Some clarifications are nedeed.
Additional comments: The paper has been improved. However, there are still few clarifications needed.
Line 25: Replace “SFFs” with “SSFs”.
Line 37: Sometimes, throughout the manuscript, the term “bycatch” is used (also in Discussion Line 295); however in the Results section, the terms used are “landed species” and “discarded species”. You should clarify this because “bycatch” and “discards” are not synonymous.
Line 72: Please, remove the excess “the”.
Line 104-113: Review the aims of the study. “iii) to widen the range of commercial species to be caught in comparison to the traditional pots used in the area;”. A proper comparison of the captured species by each type of pot is not performed in this study.
I propose something similar to “Based on these considerations, a pilot study was devised to test the effectiveness of a fully collapsible pot design in comparison of a traditional set net under commercial conditions; in terms of i) catching commercial species, iii) reducing discards, and iii) other aspects related to use and handling.”
Line 112: Replace with “these areas”.
Line 117-118: You say previously that the study is conducted in three areas. So, you could add the name of the three areas there: “and involved three coastal areas (Marina di Ravenna, Portonovo and Senigallia), where depth ranges from 5 to 19 m (Figure 1).”
Also, you should move the sentence “Specifications of bottom type and average depth of the three sites are listed in Table 1.” behind the sentence “and involved three coastal areas (Marina di Ravenna, Portonovo and Senigallia), where depth ranges from 5 to 19 m (Figure 1).”
Table 1: The SD in depth should be added.
Line 146: Please, move the reference at the end of the sentence.
Supplementary Table 1 y 2: These tables have greatly improved their clarity. Only in the column headings you use “Big CPUEW” and “Small CPUEW” instead of “LP CPUEw” and “SP CPUEw”.
Also, I insist again on a point already discussed in the previous review. You say (line 157-160) “The total catch for each gear was thus divided into a landed catch (species with commercial value, not necessarily target species) and discards, i.e. species discarded for different reasons (invertebrates and fish species with no commercial value, commercial individuals under the MCRS or in poor conditions)”. So, why do you include the commercial individuals under the MCRS as landed or commercial (i.e., marketable) catch? They should be included as discarded species in Table 2. And, are this fraction (i.e., commercial individuals under the MCRS) included as landed catch in all analyzes carried out on the manuscript? If so, the results obtained are not entirely correct. I understand that, at this point, to modify this is a great effort, but I encourage the authors to carry out this modification since it would give more reliability and robustness to the results.
Line 208-210: “Other landed species caught by the pots were S. umbra, Conger conger (LP) and S. umbra (SP) at Marina di Ravenna and Dentex dentex (SP) at Portonovo (Supplementary Table 1)”. Please, review this sentence. S. umbra at Marina di Ravenna is named twice.
Line 230-231: Pots will be a valid alternative to GTRs, just if pots are equally or more economically profitable. The profit of the fishery depends mainly on the catches of S. officinalis (the most abundant capture and with highest price (I suppose)). Therefore, this is a very important point: “The catch comparison curves (Figure 6 and Table 3) demonstrate that for S. officinalis, pots (both dimensions) were more efficient than GTRs”. However, it is not clear enough if this higher efficiency is limited only to certain sizes. Could you clarify this? Maybe an additional analysis could be needed with the CPUEw of cuttlefish.
Line 252-259: The main results obtained in this study should be added in this paragraph. Also, you could discuss about the economic efficiency of the pots vs GTRs.
Line 262: Replace “weigth” with “weight”.
Line 260-263: This part is referred to discarded fraction. So, you should move to the paragraph that begins in Line 275; specifically, when you talk about the obligation to land. Also, you could add the percentages obtained in this study.
Line 295: Review the use of the term “bycatch”.


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: MESOZOOPLANKTON GRAZING MINIMALLY IMPACTS PHYTOPLANKTON ABUNDANCE DURING SPRING IN THE WESTERN NORTH ATLANTIC
Review round: 1
Reviewer: 1

Output:
Basic reporting: The study is generally well performed and the ms well structured and  written by native English speakers (as presumed by names and affiliations) in clear, professional language. As a non-native English speaker I cannot comment further on that. The Introduction to the ms provides adequate background information and puts the problem under investigation in a proper reference framework; important and relevant bibliographic references are provided in support of authors’ statements. There are, however, a few instances where the message may not be completely clear. For instance:
I found a bit confusing the following statement (lines 64 – 66): “Thus microzooplankton are thought to respond faster than mesozooplankton to increasing phytoplankton availability leading to bloom events (Sherr & Sherr 2009).”. The first part of the sentence is clear-cut, but I do not see how the second part is logically derived from the first. I mean that microzooplankton grazing is usually considered a first order regulator of phytoplankton population-community dynamics, preventing the formation of blooms on most occasions (i.e., blooms happen when grazing control fails; for instance Irigoien, Flynn & Harris 2005, JPR 27: 313-321, Beherenfeldt 2010, Ecology 91: 977–989; Beherenfeldt & Boss 2018, Glob Change Biol 24: 55–77; ). As it stands, the second part of the sentence seems contradictory with such understanding. Could authors please rephrase?
Also, the following statement seems unclear to me, or not completely correct (lines 66 – 68) “While grazing pressure of mesozooplankton on oceanic primary production (PP) can occasionally be comparable to that of microzooplankton, in productive oceanic regions microzooplankton tend to exert a higher grazing pressure than mesozooplankton”. Microzooplankton grazing rates tend to be higher than mesozoo’s on both high and low productive ecosystems, according to the series of meta-analyses and synthesis by Calbet and collaborators (Calbet & Landry 2004, Limnol. Oceanogr., 49: 51–57; Calbet 2001, Limnol. Oceanogr., 46: 1824–1830; Calbet 2008, ICES J Mar Sci 65: 325– 331; Calbet & Saiz 2005, Aquat Microbial Ecol 38: 157– 167).

Figures are all relevant and clearly presented. One suggestion would be to provide an extra figure (maybe as supplementary matter) to show the results from the dilution experiments, i.e., net growth vs %WSW. That would aid readers to follow better some of the deductions and interpretations of results from Sts 2 & 4. No reference is made to raw data availability.
Experimental design: A general problem I find in relation to the approach to derive grazing estimates is that when both meso and microzooplankton are present in the bottles, grazing rates by both grazer communities are not additive due to interactions among the many components, including intra-guild predation. Strictly speaking that is a problem inherent to the seawater dilution method where complex micro to pico-sized communities are manipulated. Adding mesozooplankton enhances the chances and ranges of such interactions, and thus increases the difficulties in the interpretation of the experimental designs. In the Discussion section authors properly address some of this issues.

Besides the above, the methods followed are currently largely accepted. I highlight positive issues like the correction performed to take into account photo-acclimation effects. Indeed, the validity of the whole approach based on chlorophyll would be seriously compromised without that improvement.

It is unclear the reason why C. hyperboreus was selected as representative copepod of S1 considering that the most abundant was C. finmarchicus. Could authods justify their choice?.

The description of the methods is generally fine, but there are several aspects that are not well covered, or simply deserve further detail. Main issues follow:
- filtered (S1) or unfiltered (S2) seawater was used to keep zooplankton for later sorting. why not a consistent protocol was followed?. Please justify.

- Use of a 220 um mesh net clearly underestimates abundance of small copepod species (Paracalanus spp. and Oithona spp.) and also naupliar stages of larger ones (Calanus spp.). Those issues could be somehow accounted for when discussing the field implications of current results.

-How much unscreened water was added to the experimental bottles (copepods treatment) at S4?. Did that resulted in approximately realistic copepods concentrations within the bottles?.

- It is not stated in the methods whether different zooplankton concentration treatments were also considered at S4?. Please specify.

-Please indicate the volume of water analysed in order to characterize particles size spectra.
Validity of the findings: No comment
Additional comments: Other issues:

Results
It is stated that “No microzooplankton grazing was detected at S2” (lines 292-293). Could authors please be more specific?. One understands that net growth measured at 100%WSW was exactly the same than that measured at 20%WSW. If actual results were shown this doubt would be easily evacuated.

It is important that the number (or concentration) and relative composition of the mesozooplankton in the experimental bottles is also reported (for S4). A priori one may expect relative composition inside the bottles to be identical to that in the field community, but due to all necessary manipulations for the preparation and addition of the sample to the bottles, the experimental community may deviate significantly from the field one.
Related to the above: In line 300 text reads “Inclusions of mesozooplankton at in situ abundances and… “. Which indications exist that mesozoo concentration inside the bottles properly represented in situ abundance?. Could authors present evidence on that?

I assume particles size spectra shown in Fig. 6 are averages from replicate samples/ bottles. Please specify, and provide some error index for between-replicates variability (could be just a quote in the text, or in the legend to thefigure).

It is stated that “… all particles in the size fraction 15-60 μm were removed by mesozooplankton grazers at all stations over the course of the 24 h incubations” (lines 338-340). However, Fig. 6 clearly shows the presence of particles >15 um in all stations and under all conditions tested. Maybe I am not properly interpreting these results; could authors revise and/ or explain better?.

Lines 348-350: “Blank controls contained a negligible amount of particles and replicate samples revealed a coefficient of variation of 1%.”. Please specify if that CV refers to the total particle count from replicate FSW samples.

Discussion

At St. 4 particles abundances seem to be slightly higher in Microzoo treatment compared to the initial condition. As the ordinates axis is log10-scaled, at certain size classes the difference may be substantial. Any clue which may be the reason for that?. What about population growth?

I do not completely agree with the statements in lines 366-369. And consequently with the proposed shifting balance between micro and mesozoo grazing. Justification: Grazing by microzoo at St 4 was undetermined, not zero. Those results are obscured by the fact that in that case no photo-acclimation correction could be made. The unexpected pattern (lower growth at 20%WSW) could be related to that. In any case, if the basic assumptions of the method were not met, that does not necessarily leads to the conclusion that grazing was zero. This issue is revisited later in the Discussion (line 474), and this second time the interpretation is different and more rigorous (and correct, in my opinion).

Lines 422-423: “The low impact of grazing by copepods on phytoplankton observed here suggests that copepods likely relied on other food sources.” That is not necessarily so. Copepods may obtain 100% of their required daily ration from phytoplankton and still use a small fraction of primary production. It is a matter of copepods field biomass concentration and biomass-specific energetic requirements: copepods are most of the time too few per unit ocean volume to induce a higher impact on primary production (please note: indirectly this is correctly discussed on previous paragraphs that deal with potential aggregation effects). Of course, on top of that, copepods also consume other non-phytoplankton items. We completely agree on that. Please rephrase.

Consider that copepod densities used in the bottles of S2 are very likely unrealistically high (10000 – 40000 C. finmarchicus m-3). That may have helped to get a clearer signal but at the same time may have induced an underestimation of activity (grazing) rates. Probably a similar consideration is valid for C. hyperboreus at S1?.

Statement in lines 437-439: Yes, I generally agree with the speculation that copepods likely preyed upon both microzoo and phytoplankton to some extent. But consider that meta analyses suggest that there is an overall trend for copepods to select microzoo (actually ciliates) over phytoplankton (Calbet & Saiz, op. cit).
448-449: statement “… the situation would be similar in situ, as feeding on larger cells would be limited by the low in situ abundance of larger particles”. Well, not necessarily so. Limitation is more likely during the experiments due to very high copepods concentrations used and limited bottle volume.

Lines 495-516 (par on light effects): General agreement. But, for the lack of light effects on grazing consider the following: if copepods grazing is developed mostly during night ours and only marginally during daytime, then light intensity during non-feeding hours may irrelevant and not lead to a light effect on grazing rates (under the assumption than the photoperiod remains constant, as it is the case here).


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: MESOZOOPLANKTON GRAZING MINIMALLY IMPACTS PHYTOPLANKTON ABUNDANCE DURING SPRING IN THE WESTERN NORTH ATLANTIC
Review round: 1
Reviewer: 2

Output:
Basic reporting: see summary text
Experimental design: see summary text
Validity of the findings: see summary text
Additional comments: This manuscript present results from three sampling in the North Atlantic during the spring bloom. The stations are widely separated and range from 4-15 °C, and obviously two very different water masses. S1 and 2 are arctic with large Calanus dominant, S4 är temperate with a dominance of smaller copepods. At each station the microzooplankton and mesozooplankton herbivorous grazing is measured or inferred through the dilution technique. The particle size spectrum is measured at the three stations, before and after incubation with copepods.

The experiments are OK and well presented, but only 2-3 replicates are set up. The addition of copepods is very high, 5 C. hyperboreus and up to 40 C. finmarchicus added to 1.2 L bottles is very crowded. Hence clearance rates are very low for both species. Certainly, they do not perform anywhere near natural under these conditions. Therefore, any attempt to measure grazing potential (either as %PP or % phytoplankton biomass) will likely be underestimated. Indeed, the authors discuss this at ambient abundances and conclude that the grazing should be dominated by microzooplankton at S1. This should probably be so at S2 and S4, but here they did not get a negative slope and thus could not estimate the microzooplankton grazing.

At S2 and S4 there should be strong cascading effects of copepods feeding on the microzooplankton. The authors should take this into account. C. hyperboreus probably do not feed on ciliates, but C. finmarchicus and the smaller copepod assemblage would certainly change the ciliate concentrations when added at these high abundances. It would be interesting to note this in the analyses and in the discussion.

The chlorophyll was analysed by extraction, but indicate what pore size filters were used. This important since the flow cytometer also collect particles smaller than 0.7 µm, the standard nominal pore size of GF/F filters. The dilution series were prepared by 0.45 µm screened water, were these filters also used for the chla determinations?

The grazing is inferred from the slope in Fig 4, but the regression tests are wrong. The df in line 302 and 307 regressions is 1, not 6. Significance seems OK.

The two-way ANOVA should be presented with all relevant df, SS, MS, F and p-values. The discussion of the results from the ANOVA (lines 317-333) is unclear, but will be clearer if the table is presented.

What was the absolute abundance of the copepods? (Fig 3). Please give this along with percent composition.

Fig 6. Are there any replicates for these data? Error bars for the measures? You should also provide statistical tests for the differences between treatments, e.g. Kolmogorov-Smirnov tests

Table 1. The ABC for the different bottles are not shown as is indicated in the legend, please correct

Line 530 This conclusion (no preferential feeding …) is not well supported by data. Explain better or remove.


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: MESOZOOPLANKTON GRAZING MINIMALLY IMPACTS PHYTOPLANKTON ABUNDANCE DURING SPRING IN THE WESTERN NORTH ATLANTIC
Review round: 1
Reviewer: 3

Output:
Basic reporting: The manuscript reads very well and generally speaking the flow and contextualization is great. The objectives are clearly stated. It is very nice to see studies on micro-zooplankton grazing rates combined with meso-zooplankton, as this is often ignored and the former, as shown here, can be quite significant. However, because the micro-zooplankton grazing measurement is often ignored, many readers will not be familiar with the dilution method, even less so with the modified method used here. Thus, below are some general and specific suggestions to improve the flow and make it easier for readers to understand and replicate the study.

General issues

1) I would suggest a graphical representation to visualize the experimental design for the grazing experiments. The method is quite complex with a specific concentration gradient of meso-zooplankton at each site crossed with different nutrient conditions, micro-zooplankton grazing via dilution experiments, and also different light intensity in one of the sites (S4). Thus, it may be difficult for many readers to understand how the micro- and meso-zooplankton grazing effect was calculated and compared, especially for those that are not familiar with the modified dilution method. I suggest including a graphical image of the overall experimental design to measure the micro- and meso-zooplankton grazing for S1, S2 and S4. This would undoubtedly help readers understand and provide a clear vision for anyone who wishes to replicate or build on what was done.

2) The grazer effects and comparison of the differences among micro- and meso-zooplankton effects presented in Table 1 (or from the other results) is not straightforward. It might help to calculate the overall effect of each grazer type on % primary production, for example (see more on this below in “validity of findings”).

3) I suggest to highlight more clearly in the results that S1 had the most grazing by micro-zooplankton, that S2 had the largest impact of copepods on particle size distribution but no observable microzooplankton grazing, and that S4 had no significant grazing effect by either meso-zooplankton or micro-zooplankton.

Specific issues:

L 241-242: Many readers will not be too familiar with the modified method for calculating micro-zooplankton grazing rates used here. In this context, the difference between k (net phytoplankton growth rate) and µ (in-situ specific phytoplankton growth rate) and their importance for the method can be better explained.  Moreover, I suggest calling µ the “total phytoplankton growth rate”, which is more descriptive than “specific growth” (if I understood correctly, µ is the total growth rate = grazing + k). This also goes for Table 1.

L 242: “the sum of the grazing rate and the mean net growth rate of the non-amended <200 μm WSW bottles”. I understand that the grazing rate here is the micro-zooplankton grazing rate (i.e., “g”). If so, please make this clear by adding a parenthesis “(g)” after “grazing rate”.  Also, why is the µ in Table 1 not the sum of (g + k NoN)? If I understand correctly, mean net growth rate of the non-amended <200 μm WSW bottles is k NoN in Table 1. Thus, µ should be the sum of g + k NoN (=0.40, not 0.39), right?

L 346-348: Do you mean for all sites? If so, please state clearly.

L 361: “… observed shifting balances …” shifting in what context? “Shifting balances” here may be replaced with “We observed significant variability in the micro- and mesozooplankton grazing impact” to avoid the possible temporal implications that may be associated to “shifting”.

L 368: Why is this remarkable? Would light limitation not be expected to be a major loss factor here? Please explain in a few words.

Table 1: I believe A, B, and C are missing in the column for k Cop.

Figures 4 and 6: There is data for all of the treatment copepod concentrations in Figure 4 for S1 and S2 (i.e., 3 per site) but not in Figure 6? Please add the data for these different copepod abundances or explain why they are not included.
Experimental design: The topic is within the scope of journal and there is a well defined research question. The authors did a good job in describing the relevance of this work and highlighting the knowledge gaps. The methods are sound but could be explained in a more straightforward fashion, especially with regard to why the nutrient additions were made and the logic for using nutrient added or non-added bottles while calculating grazing impacts. Doing so would improve future efforts in replicating the methods and approach used here. Please find below some suggestions for improving the manuscript in this context.

General comments

1) It is not straightforward why the micro-zooplankton grazing rates are based on phytoplankton growth rates from the bottles with nutrient additions while the net phytoplankton growth is calculated from non-amended bottles. Please explain the reason for this and see the specific comments below.  This would also help to understand and interpret results presented in Table 1.

2) Given the difference in copepod size in S1 and S2, it would be nice to provide the length and biomass of the species used. That way the reader and the authors can compare the mass-specific top down effects.

3) The in-situ grazer density (i.e., natural density during sampling) is not reported, which makes it difficult to compare the experimental grazer density with that of the natural grazer density at the time of the experiment. The authors mention that it was not possible to estimate this density but it would great to include some estimates from other studies to provide the range that might be expected in nature.

Specific comments

L 235: Why is the microzooplankton grazing rate (g) calculated from phytoplankton growth rates (k) obtained from the bottles with nutrient addition only (k +N)? I believe many readers, including myself (!) would like to know the justification for this.

L 242: Similar to the comment above for L 235, please explain why µ is calculated using the net phytoplankton growth from non-amended bottles? It is interesting that g is calculated from k + N, while µ is calculated from k NoN and the conceptual reason for this should be clear.

L 341-345: What happened to the data from the bottles with different copepod densities for S1 and S2? Please explain why there are 3 different copepod concentrations in the grazing experiment (S1 and S2) but only 1 copepod concentration for S1 and 2 copepod concentrations for S2 in Figure 6?
Validity of the findings: Overall, the conclusions and their relevance to the objectives are clearly stated and are mostly justified by the results presented. Combining no-grazer controls in addition to different grazer concentrations adds to the strength of the meso-zooplankton grazing experiments. One key limitation might be the lack of a universal metric to compare the effect of micro- versus meso-zooplankton on phytoplankton growth (e.g., % reduction in PP). There are a few more suggestions for improving the manuscript in terms of strengthening the conclusions provided below.

1) Currently, the relative effect of micro-zooplankton is not directly compared to the effect of meso-zooplankton on phytoplankton. Thus, the justification for concluding that micro-zooplankton had a stronger effect compared to meso-zoopankton is not straightforward. I suggest the authors to make this comparison clearly before proposing general conclusions between the grazing impact of micro- vs. meso-zooplankton.  For example at S1, did microzooplankton graze more phytoplankton or did copepods? L 292 says that 97% of primary production was grazed by micro-zooplankton, but this is for the dilution experiment only. How does this compare to what was grazed down by copepods at the same site (S1)? Based on the k copepod values in Table 1, can we conclude that the highest copepod density in S1 (k Cop= - 0.40 relative to the k NoN = 0.02; a difference of 0.42 d-1) consumed more phytoplankton than micro-zooplankton (g = 0.38)? It is not clear if this kind of comparison is possible. Either way, the quantitative effect (e.g., % reduction in phytoplankton biomass or growth) among the micro- vs. meso-zooplankton should be made clear for S1 and also estimated for S2 and S4.  If this is not possible, please explain why not.

2) The conclusions regarding the grazing effect of copepods on micro-zooplankton (e.g., Line 530) may not be entirely justified because there is no specific information on this in the results. Please either re-formulate these conclusions, or clearly state in the results what data there is showing the grazing of copepods on micro-zooplankton.  More on this in the specific comments below.

3) It would add significantly if the authors could include the mass-specific CR data (i.e., mg prey mg copepod-1 day-1).  The prey here could be in terms of chl-a, or biovolume. This would enable others to compare among previously published grazing rates and give an idea of the grazing pressure to those more familiar with CR. See specific comments below for more on this.

4) It is very nice how the authors showed the effect of copepods on the prey size spectrum. Would it be possible to estimate size specific grazing rates? If the copepods ingested large sized phytoplankton, perhaps it would be interesting to estimate the CR based on small vs. large prey.

5) Given the latitudinal gradient of the study, it might be interesting to speculate about the geographical differences in the observed results. While the opportunistic sampling indeed limits such generalizations (as recognized by the authors), it is interesting that the colder, more mixed water (typically where we might expect stronger copepod grazing pressure on phytoplankton) had the strongest micro-zooplankton grazing impact. What do the authors think about this? Just a result of stochastic mechanisms? A few words on this in the conclusion would add to the manuscript.

6) Would meso-zooplankton grazers including copepods increase at night due to vertical migration? By what factor would this increase the grazer abundance? Would be interesting provide a few lines on the possible shifts in copepod grazing pressure due to vertical migration.

Specific comments

L 249: It is difficult to compare per capita CR among different sized grazers. Larger grazers will have a higher CR. Thus, it is more appropriate and informative to compare mass specific CR. That way one may compare the top down effect in different sites and other workers, including models, may compare with your results. If possible, please provide the mass-specific CR estimated from S1 and S2 for the different copepod species used.

L 317: Is it not possible to also calculate the mass-specific CR from S4? If the number and size of the copepods and other meso-zooplankton in the grazing bottle is known, than it would be possible and useful to calculate mass-specific CR. By doing this also for the S1 and S2 experiments, then you could compare the mass-specific CR across all sites for a more integrative view on mesozooplankton grazing.  I believe this would make the study stronger and more informative.

L 361-363: Please see the general comment #1 under the Validity section.

L 366-368: Could the % reduction of phytoplankton growth due to meso-zooplankton be estimated for S2 and S4?

L 384-395: This is great, but please also add some published copepod densities reported for this region to back up the statements made. For example, what is the common recorded copepod abundance and the exceptionally high “peak” abundances in the NW or mid-Atlantic? These numbers would make the point much stronger and add to the empirical context of the presented results.

L 419-421: Having a clearly stated effect of copepods on PP (% reduction) from this study would help put these results in context.

L 452-454: Unfortunately, this statement, as it is currently written, is not justified by the results. If I understand correctly, there is no information in the current study regarding the grazing impact of copepods on micro-zooplankton. Also, is the 15% reduction in PP a result from this study? If so, from which site? Please make this information more straightforward.

L 454: Which size class exactly? Please specify.

L 530: Please indicate where in the results this information about the degree of micro-zooplankton grazed by copepods is presented.
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: MESOZOOPLANKTON GRAZING MINIMALLY IMPACTS PHYTOPLANKTON ABUNDANCE DURING SPRING IN THE WESTERN NORTH ATLANTIC
Review round: 2
Reviewer: 1

Output:
Basic reporting: No comments
Experimental design: No further comments
Validity of the findings: No comments:
Additional comments: Answers to comments on the first round of review show an effort to consider all issues raised by different reviewers. That means inclusion of extra information required (for instance regarding the methods) as well as taking care of interpretation issues or clarifications requested. I find the ms is in better shape now.
A single further comment (I must have overlooked this on the first round) is that I could not find a reference in the Methods to actual dates when stations were occupied and experiments run. At the end of the Introduction it is stated that this investigation was carried out during the NAAMES spring campaign. It is good to mention that. However, oceanographic condition during for example end of March may be rather different from that in early June, and both correspond to Spring. Similarly, information on the year the oceanographic cruise was set is in order, as year to year environmental variability could be significant. I think provision of more precise information on the actual year and dates is valuable for reference and comparison for future investigations, and will thus enhance the utility of current data and results. I thus strongly recommend the addition of that 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: MESOZOOPLANKTON GRAZING MINIMALLY IMPACTS PHYTOPLANKTON ABUNDANCE DURING SPRING IN THE WESTERN NORTH ATLANTIC
Review round: 2
Reviewer: 2

Output:
Basic reporting: The authors did a good job at revising their manuscript and I have no further comments or suggestions.
Experimental design: The authors did a good job at revising their manuscript and I have no further comments or suggestions.
Validity of the findings: The authors did a good job at revising their manuscript and I have no further comments or suggestions.
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 SWITCH FROM ONE SUBSTANCE-OF-ABUSE TO ANOTHER: ILLICIT DRUG SUBSTITUTION BEHAVIORS IN A SAMPLE OF HIGH-RISK DRUG USERS
Review round: 1
Reviewer: 1

Output:
Basic reporting: The language becomes casual in parts and there are a number of erroes in punctuation and sentence structure.
There was sufficient background and context.
The article could be restructured to flow more easily. Some of the tables could be substitued with graphs which might make it easire to understand.
Experimental design: The research question is not well defined.The methods are not described with sufficient detail to  be confident that the investigation has been performed with a high technical standard, or indeed that it could be replicated.
Validity of the findings: There is not enough data in the methods on participant recruitment and what was assessed (frequency of use, period over which this was discussed) to fully appreciate the results.
Additional comments: What does drug substitution behaviour tell us about the deficiencies in our drug treatment service. Illicit drug substitution behaviours in a sample of high-risk  drug users.
The title is misleading. This study describes substance use in a cohort engaged in various treatments for SUD. I don't think it can really identify deficiencies in drug treatment services.
The paragraph “Purpose of the study” is a bit confusing. The difference between aims and objectives is not helpful.  The sentence “Data provided by treated individuals on current and past drug use was used to ascertain the prevalence of substitutions” might be better placed in the methods section. It is perhaps more typical to describe aims then clarify the  primary and secondary outcomes n the methods section.
There are a number of problems with the methods. This current study is reported to be part of a larger study. This is not explained in sufficient detail, or indeed why the current study is being presented. The date range of the study is not provided.

The nature of the population examined is a bit unclear. There needs to be greater detail in the definition of a high risk drug user, rather than just the citation.
The paper could be improved by a number of clarifications about the population under study.
(1)	The source of participants could be clarified: Detoxification and Opioid Agonist Treatment (OAT) are not the same. Line 177 describes physical detoxification in hospital, ambulatory units or in methadone centres where participants are receiving OAT. Line 194 talk about drug users undergoing detoxification. Are participants being “detoxed” in methadone centres ? The term “Physical detoxicfication” is unfamiliar and requires clarification.
(2)	The stage at treatment at which participants are recruited should be clarified.
(3)	A description of how participants were recruited to describe potential sources of bias from recruitment strategies would be useful. This would help identify to what extent the participant population reflects the populations attending these treatment centres.
(4)	There needs to be greater detail of the questions asked of the participants. Was a standardised questionnaire used. Over what period of time prior to interview was drug use ascertained? Is recall bias therefore a problem? Is there any identification of frequency of use? Perhaps a sample questionnaire could be appended to the paper.
(5)	Were the doses of OAT identified as inadequate OAT dose is a significant contributor to continuing opioid use.
The results are quite difficult to read and a bit disjointed. The order of presentation is a bit unusual. It is conventional to describe the demographic characteristics of the sample, and analysis done of that group, and then present the subgroup description and analysis sequentially. Presenting the substitution data graphically (pie chart) might be visually more appealing than a table, and easier to understand.
There are a number of typographical and grammatical errors throughout the manuscript, and the language can become a bit colloquial (eg NIMBY).


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: MECHANISMS BEHIND BOTTOM-UP EFFECTS: EUTROPHICATION INCREASES FECUNDITY BY SHORTENING THE INTERSPAWNING INTERVAL IN STICKLEBACK
Review round: 1
Reviewer: 1

Output:
Basic reporting: “Mechanisms behind bottom-up effects: 1 Eutrophication increases fecundity by shortening the interspawning interval in sticklebacks”  is a informative study about the potential mechanisms of  bottom up effects of eutrophication on population dynamics in sticklebacks.
The article is well structured, while there is no figure in the publication the included tables are easy to read and informative enough.
The authors deliver a good introduction reporting the different ways eutrophication can interact with community composition and dynamics of the ecosystem.
Experimental design: The authors conducted a lab experiment where they find that eutrophied conditions in the field with high amount of algae and food can favour shorter inter clutch intervals and thus lead to a higher female lifetime fecundity.

- The study itself uses 2 treatments, high algae + more food, and low algae + little food. The authors discuss that the observed  the effects on lifetime fecundity could be due to the “protection” by algae (l194ff) or from an increased food availability (l180ff). While the two chosen treatments reflect the conditions in the field, it would have been great to include the other two combinations as well, to be able to discern the relative contribution of both. I can see that the workload is already very high with just the  two treatments, but should maybe be mentioned in the discussion as a “blue print” for further studies. Or it could be indirectly addressed by trying to compare the observed change in interspawing interval to changes observed from experiments with varying feeding levels. (see general questions below)
Validity of the findings: The findings are valid, the results are not overly interpreted and shortcomings due to the experimental design are addressed.
Additional comments: Here are some general comments, questions, and minor corrections  which I think would improve the manuscript:

- How does the reduction of the inter-spawning interval in this study compare to previous studies with varying food availability? I think this information could be useful to discuss how much of the effect could be due to food compared  to stress relief.

- l 223, maybe add “…and an increased number of nests in dense vegetation”? I guess that is more or equally important than hatching success, especially  since Stickleback males do not take as many eggs as possible, but “close” their nest after a while, hence females would be limited by nests to place their clutches in this limitation could be removed in eutrophied areas.

- One topic i was missing is how does predation risk change in eutrophied areas? do females or offspring get eaten more or less in eutrophied areas? While trying to scout the literature I came across this paper : Gagnon et al 2017 (Shifts in coastal fish communities: Is eutrophication always beneficial for sticklebacks? ) where the authors claim that eutrophication/filamentous algae are actually negatively correlated with stickleback abundance in the field, partly because there is less cover against predation in these areas. While I think their stats are a bit weak, it would maybe be good to discuss your results in perspective with theirs.

l37: shouldn’t it be rather: “the enrichment of ecosystems with nutrients?”

l56: ..”such as climate change” is it through  warming or the predicted precipitation/freshwater inflow. It would be nice to be more specific here.

l61. “…calling for more research into the topic.“

l96 maybe specify:” the larvae of piscivores” to be clear that they are not eating their own.

l110 the description of the location could be more precise, at the moment it leads me to somewhere in the baltic outside of Helsinki.

l128 “thank” should be “tank”

l131. added nutrients to one but not the other treatment… Could this also have influenced the results? I cannot see any obvious connection, given that sticklebacks obviously do not feed on nutrients, but I think it deserves a mention…


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: MECHANISMS BEHIND BOTTOM-UP EFFECTS: EUTROPHICATION INCREASES FECUNDITY BY SHORTENING THE INTERSPAWNING INTERVAL IN STICKLEBACK
Review round: 1
Reviewer: 2

Output:
Basic reporting: 'no comment'
Experimental design: l.102-106 - The aim should be more clearly presented. Remove "and an increased density of filamentous algae and prey abundance" (l.102) and consider this version: "We investigated if eutrophication increases female fecundity during the spawning season." as you explain the details of your treatment in the next sentence.

l.110 - please provide more exact coordinates

l.114 - was Standard or Total body length measured?

l.132-133 - please add information on what you actually added (nitrates and phosphates?)

l.136 - this is the only spot, where the "low" treatment is described as a control. For the overall perspective, consider making it more obvious throughout the text.
l.138 - fishes do not have cloaca, use urogenital papilla instead
l.157 - The raw data seems consistent with little intra-individual variation, so ANOVA on averaged values seems appropriate. I just wonder why the Authors did not use mixed-effects models with female ID as random factor to gain statistical power in their analyses and avoid losing the information on among-clutch variation.

l.157 - it seems worth to be more exact on the sample size here. I found that 20 females per treatment were analysed only after checking the original data file. Consider writing something like: "We used 20 females per each treatment". Under replicates, one can imagine individual spawning events too, but your study was much more robust, so don't be afraid to write that down. Also because of this strange wording, I wonder whether there was more females used in the experiment that did not reproduce. If so, this should be explained in the text along with their respective number in each treatment.
Validity of the findings: l.174 - I suggest to change "Nutrient enrichment and a high density of filamentous algae" to "Our high algae density treatment" as the manipulation was complex and also included adding invertebrate prey.
l.180-193 - Is there any information on what is the number of clutches in the natural population?
l.182 - I would rephrase "The enrichment with nutrients maintained high algae growth conditions across the season, which is known to" with simply "High density of algae" as it seems that algae relative growth was lower in High algae treatment compared to the low (Table 2 - (69.50-50)/50 vs. (175.96-150)/150).
Additional comments: The study by Anne Saarinen and Ulrika Candolin presents an interesting experimental approach to test the effect of eutrophication on female reproduction in three-spined stickleback. They manipulate amount of filamentous algae, number of invertebrates in the tank, and concentration of nutrients on tank inflow, to create high and low treatment. They find that despite the clutch and egg size remain similar between the two treatments, the high algae sticklebacks are able to spawn more frequently. The Authors then put their findings into the context of population dynamics under anthropogenic influence.
The manuscript reads well overall and is clearly structured.  The aims, methods and results are concise and sufficiently detailed. Although the experimental design consists of a multivariate manipulation (higher amount of algae, adding Gammarus sp., and nutrients), the use of "High algae density" is a reasonable umbrella label and seems carefully handled and described. Data analysis was performed appropriately and the main outcomes are sound. Discussion provides reader with enough context of the current research on eutrophication effects on stickleback populations, and also offers potential explanations for the observed pattern. The study therefore suits publication in PeerJ and I suggest minor changes in addition to the comments provided in the Review sections above (1.-3.):

l.37 - maybe change to "with" in "of nutrients"
l.80 - remove "to" from "do not to"
l.210 - reword to "Whether this contributes to the growth of stickleback populations..."
l.222 - "hatching success of males" reads inappropriately and I would talk about increased recruitment or even nesting success.
Tables 1 & 2 - Label treatments as "High algae" and "Low algae". "High density" confuses readers.


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: MECHANISMS BEHIND BOTTOM-UP EFFECTS: EUTROPHICATION INCREASES FECUNDITY BY SHORTENING THE INTERSPAWNING INTERVAL IN STICKLEBACK
Review round: 1
Reviewer: 3

Output:
Basic reporting: The manuscript by Saarinen and Candolin aims to test the effects of the widespread phenomenon of eutrophication (as the anthropogenic increase of nutrients) on the life-cycle of three-spined sticklebacks, particularly focusing on its effects on female reproductive traits. Using a laboratory aquarium experiment, the results showed that eutrophication leads to higher lifetime fecundity of female stickleback either by altering the habitat structure (providing more shelter) and/or increasing the productivity of the system (having higher prey availability). The mechanism behind higher lifetime fecundity was due to an increase in the number of times the females spawned (via reducing the time between spawning events), rather than changes in clutch size, egg size or the invested energy to reproduction (based on changes in body weight).
Overall, the manuscript is clearly written and understandable, it provides context and niche to the study citing relevant works, presenting relevant tables and supplying raw data that is understandable.
Experimental design: In order to test the working hypothesis (whether eutrophication increase fecundity in female sticklebacks) the authors use an experimental design in which the authors compare control tanks with a tanks subjected to a eutrophication scenario by: (1) adding more algae, (2) adding more prey, (3) adding more nutrients. Despite all three variables are likely to come together during eutrophication, this multiple simultaneous manipulation makes a bit difficult to decipher the mechanism that affects fecundity in fish. A longer term incubation before the fish addition just with nutrient additions and equal initial conditions of algae and prey would have provided a better evidence of the collinearity of these variables. Please justify the simultaneous manipulation of so many variables between control and treatment aquaria.
In addition, as the authors state in the discussion (Lines 194-219), it is difficult to separate a shelter explanation from the increasing productivity explanation of the increasing fecundity. In future studies this can be solved in by manipulating the amount of shelter with artificial substrate (e.g. plastic brush piles) independently from nutrients or food. Since the mechanisms of the effects of eutrophication are difficult to disentangle, I would tone down all the statements of highlighting the impact of this study in understanding the mechanisms by which eutrophication affect fish populations (Lines 16-17, 49-51). Additionally, since it is not clear that productivity explained this pattern instead of shelter, I would advise not to use the bottom-up (productivity) effects but more eutrophication as a mediator of both productivity and habitat changes, so please reconsider to change the title of the article either adding habitat changes or removing bottom-up effects.
Validity of the findings: The implications of these results are important and relevant, but I would be cautious when upscaling these results to patterns in the whole population or even more to the community and ecosystem level. This study shows the effects of potential eutrophication only to one part of the population (females) and during a short (yet important) period of their life histories. How eutrophication affects the males, the actual mating success and the survival and growth of the sticklebacks during ontogeny is yet unclear based on this study and may affect the outcome of population dynamics. Yet, the mechanism here presented may explain (to some extent) the trends found in natural populations (Candolin and Voigt, 2020).
Additional comments: Specific comments
Abstract
Line 24-25: the results of more offspring emerging I habitats with denser algal growth are not presented in the study. Please make it clear that this knowledge comes from other studies or delete it.
Introduction
Lines 85-101: speculative sentences that would fit more the discussion than the introduction.
Materials and methods
Line 110-111: Motivate why using the temperature and salinity conditions (based on natural conditions?).
Line 113-114: Why weighting the females after the first spawning event and not before as well =?
Line 115-116: Could you explain how the tanks where distributed spatially. Did the same water flow through the treatment aquaria?
Line 117: I assume that the 0.2 mm net clogged with algal growth during the experiment, especially in the treatment enclosures. Did you do anything about it?

Line 118-121: Can you describe the spatial design within each enclosure?
Line 132-133: Which type of nitrogen and phosphorous were added? The addition of nitrogen and phosphorous are far away from natural proportions for optimal growth (Redfield ratio is 16/1 N/P molar, 7.4/1 by mass; 1.27 N/P by mass in this study ). Please specify why adding these N/P ratios and state which inorganic N and P was added (Chemical formula).
Line 161: please estate the number of the Ethical permit used.
Discussion conclusion
Line 178 Please provide reference to the statement that that population of sticklebacks dies after one breeding season. This raises the question, how representative is this strategy in sticklebacks? Is that widespread in other populations in the Baltic Sea or over the worls? Please try to add a few sentences comparing with other populations or fish species.
Line 191-193 please elaborate a bit more (add one sentence) on why is this advantageous.
References
Please be consistent when reporting references and pay special attention to the capitalization of words in titles. Only the first word of the title should be capitalized.


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: MECHANISMS BEHIND BOTTOM-UP EFFECTS: EUTROPHICATION INCREASES FECUNDITY BY SHORTENING THE INTERSPAWNING INTERVAL IN STICKLEBACK
Review round: 2
Reviewer: 1

Output:
Basic reporting: No change to my previous assessment. The study is clearly reported, the background is sufficient and gives a good overview over the topic. The raw data is available..
Experimental design: The authors conducted a lab experiment where they find that eutrophied conditions in the field with high amount of algae and food can favour shorter inter clutch intervals and thus lead to a higher female lifetime fecundity.
Validity of the findings: The findings are valid, the results are not overly interpreted and shortcomings due to the experimental design are addressed.
Additional comments: I am happy with the changes and the responses of the authors to my and also the comments of the other reviewers.

I only found two minor misspellings in the last version:
L 241 “…. to the increased prey ….”  should be “…. to the increased prey ….” (?)
L 276 remove the second “,”


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: MECHANISMS BEHIND BOTTOM-UP EFFECTS: EUTROPHICATION INCREASES FECUNDITY BY SHORTENING THE INTERSPAWNING INTERVAL IN STICKLEBACK
Review round: 2
Reviewer: 2

Output:
Basic reporting: 'no comment'
Experimental design: 'no comment'
Validity of the findings: 'no comment'
Additional comments: I think the Authors have put reasonable amount of effort to clarify their manuscript and I am happy with the changes made.


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: MECHANISMS BEHIND BOTTOM-UP EFFECTS: EUTROPHICATION INCREASES FECUNDITY BY SHORTENING THE INTERSPAWNING INTERVAL IN STICKLEBACK
Review round: 2
Reviewer: 3

Output:
Basic reporting: The authors have addressed all the comments raised succesfully. I therefore endorse the publication of the article.

Kind regards.
Experimental design: Experimental design is now clarified and supported in previous literature.
Validity of the findings: The findings are scientifically correct and implications are now toned down according to the level of evidence provided.
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: ASSESSMENT OF BIOMASS POTENTIALS OF MICROALGAL COMMUNITIES IN OPEN POND RACEWAYS USING MASS CULTIVATION
Review round: 1
Reviewer: 1

Output:
Basic reporting: 1. The error bars may be lost in Figure 1 D) and H).  Please add into these figures.
2. Some key fatty acids (i.e. EPA-C22, DHA-C24) may be lost in Figure 4. According to the microscopic photo in Figure 3 and 5, I think there are diatoms which are rich in polyunsaturated fatty acids, particularly DHA. However,  there is no fatty acids with carbon >20 in the algal communities (Fig.4). Please check carefully the results of fatty acids composition, and revise Figure 4.
Experimental design: 1. The part, applicability of microalgae a biological enhancer for soybeans, the two applications (biodiesel production and biofertilizer assay) of mass cultivation of microalgae have contradictions each other. I think it is priority to retain lipid (biodiesel) production rather than biofertilizer aspect.
2. The biofertilizer assay is also a good filed for the application of mass cultivation of microalgae, but I think it is better to have an independent and deep study.
Validity of the findings: See General comments for the author.
Additional comments: General comments:
The extent to which amplification bias affects the accuracy of microalgal community analysis is currently unknown. This study integrates a large-scale outdoor growth model with biodiversity data from a microalgal community in 2017 to estimate the long-term lipid and biomass production potential of microalgae cultured in a mass cultivation system in Korea. In addition, the application of outdoor mass cultivated microalgae to soybean plants was investigated. This paper is very rich in experimental contents, including microalgal cultivation, the analysis of genes sequencing, biodiversity, environmental parameters and nutrients, assessment of lipids content and quality, and application aspects. It is of great importance to evaluate the potential of outdoor mass cultivation of microalgae on energy production and the fertilizer industry. However, the presentation is not compact enough due to dispersed contents. Some spelling mistakes are found. The detailed comments are as follows:

Abstract:
Line 39: the biodiversity analysis of mass cultivation systems is based on high throughput sequence (HTS) in this paper, while “using metagenomics” here is not accurate.
Line 48: The seasonal changes and diversity of other organisms are the main factors affect mass cultivation of microalgae. More detailed results should be given on how these factors affect the systems. And, what are the other organisms? Other microalgae except for Desmodesums sp. or cyanobacteria and others? Line 46-48, the sentence is unclear.
Line 51-52: It is suggested to give the important data that can directly show OPR #1 enhanced the overall lipid yields.
Introduction
The whole introduction part seems to be not focused on the key points. In this part, the authors emphasize on sequencing technology, especially the primers selectivity in HTS. However, the main points in this study are effect of biodiversity in different systems on microalgal cultivation, energy production and its application. As far as l know, the THS technology is worldwide used and very common and the primers V4, V8-V9 the authors mentioned are reported previously. This paper is totally different from the paper by Bradley 2016 the authors cited. Thus, it is advised to reduce some contents on sequencing technology and give more information on factors affecting outdoor mass cultivation of microalgae.
Line 94: this cited paper was published in 2012, which was not recently published.
Line 115-116: “it had identified that conserved regions that may be best suited for amplifying hypervariable regions”. I am confused about this conclusion. The conserved regions or the conserved regions close to hypervariable regions?
Line 126-126: The V4 primer is commonly used in amplifying eukaryotic organisms. However, previous study by Bradley had been found V8-V9 was more suitable for amplifying eukaryotic organisms. The authors then use this method in the present study. Thus, these descriptions here need to be modified.

Material and methods
Line 138: here semi-continuous cultivation was carried out, while in line 153, Continuous cultivation was carried out.
Why use the groundwater with Eco-sol medium rather than domestic or unban wastewater. They contain similar nutrients level.
Line 146: CO2 was injected at 10 L min-1. What is the CO2 concentration?
Line 165-188: Miseq-based microalgae diversity, this part is too complicated. It is highly recommended to conclude the information in a table where specific primers are listed for amplification of microorganisms.
Line 179: the conserved region of V8-V9 was used for PCR amplicon?
Line 190: mock community construction. This part seems not important in this paper. As I found, this part was similar to the published paper by Bradley 2016.
Biofertilizer assays: this paper tries to evaluate the potential of outdoor mass cultivation of microalgae on energy production and the fertilizer industry. Although both aspects are showed better applicability, it seem that these two applications of mass cultivation of microalgae have contradictions each other. It is priority to retain lipid production rather than biofertilizer aspect. The biofertilizer assay is also a good filed for the application of mass cultivation of microalgae, but it is better to have an independent and deep study.
Line 243, 263-269: the fonts of equations need to be uniformed with the contents.

Results and Discussion
The structure of this part is not compact enough due to dispersed contents and results description is not concise, e.g. line 342-345,396-408. The four main contents: (a) Qualitative significance verification of HTS analysis, (b) Microalgal biodiversity of MiSeq analysis, (c) Microalgal succession during mass cultivation, (d) Microalgal and cyanobacterial compositions during mass cultivation combine as biodiversity of mass cultivation should be better.
Line 326: the prokaryotic community was analyzed using a well-known primer (specific primer needs to give here) set for 16S rRNA. The percentage of other bacteria except cyanobacteria is nor displayed. Do the bacteria have some influence on mass cultivation of microalgae?
Line 358-368: The discussions here are not closely related to the former results.
Line 365-371: this part introduces biomass and lipid productivity, and calorific value, which is again displayed later. It is better to remove this part.
Line 444-445: what are r-strategists?
Line 474-479: did nutrients starvation happen in the mass cultivation? What is the purpose of discussing on nutrients starvation?
Some spelling mistakes are found, e.g. line 433, 454, and 499.


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: ASSESSMENT OF BIOMASS POTENTIALS OF MICROALGAL COMMUNITIES IN OPEN POND RACEWAYS USING MASS CULTIVATION
Review round: 1
Reviewer: 2

Output:
Basic reporting: The authors used the metagenomics technology to combine the large-scale outdoor growth model with the biodiversity data of microalgae community, trying to estimate the lipid content and biomass production potential of microalgae in the open ponds. The data has been enough. But there are still some apparent flaws in the paper. Therefore, the paper is considered to be accepted after careful revision. The commons are as follows:
1.	I suggest that you improve the description at lines 46-54, these sentences are complicated. Please simplify the language.
2.	Rewrite the introduction. Your introduction needs more detail. The main content in INTRODUCTION is the method used in this study. So, if the authors wanted to report the findings from the view of “the new method”, please consider revising the title of this paper. Besides, please consider providing the related results of the traditional method to the algal community in the authors’ cultivation systems in order to highlight the accuracy and convenience of the new method.
But, if the authors wanted to report the findings from the perspective of the “Algal Community”, in the INTRODUCTION，they need to state the current findings about the algal community in the open ponds and the things needed to research in the future, to emphasize the novelty of the findings in this paper.
3.	In the whole text, please organize by importance and the highlights of the issues.
4.	In “Results and discussion”, It seems only to be the result description. Please consider digging the deeper reasons, such as, the relationships between the changes of the various environmental conditions and the algal community, algal lipid content, algal biodiesel quality and so on. There are not enough references for the analysis of the results. Especially for the discussion of each index result.
5.	Please consider comparing the results in this paper with those in others.
6.	In lines 358-364, the analysis of the reference can not be well compared to your study.
Experimental design: 1. The authors seemed not to clearly describe the purpose of this experiment. Please consider explaining why the authors did the research about the algal community in the open ponds.
2. For the “Biofertilizer Assays”, please consider stating the aim of each experimental setup.
Validity of the findings: NO COMMENTS.
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: ASSESSMENT OF BIOMASS POTENTIALS OF MICROALGAL COMMUNITIES IN OPEN POND RACEWAYS USING MASS CULTIVATION
Review round: 2
Reviewer: 1

Output:
Basic reporting: I feel that the abstract can be more concise.
Focus on the important results and choose the points you want to emphasize.
Experimental design: No comment.
Validity of the findings: No comment.
Additional comments: This manuscript has been improved.
I agree to publish the manuscript.


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

Input:
Title: ASSESSMENT OF BIOMASS POTENTIALS OF MICROALGAL COMMUNITIES IN OPEN POND RACEWAYS USING MASS CULTIVATION
Review round: 2
Reviewer: 2

Output:
Basic reporting: All the revisions have been well done. But the article needs to be revised by native English speakers before it can be published
Experimental design: no comment
Validity of the findings: no comment
Additional comments: All the revisions have been well done. But the article needs to be revised by native English speakers before it can be published