D. magna in Combination with M. aquaticum Inhibited the Bacterioplankton in Eutrophic Water
, Xiaocheng Wei 1, Hanyang Wang 1,2, Jiarui Li 3, Xiangqun Zheng 1, Chunxue Zhang 1,* and Bo Li 3,*Round 1
Reviewer 1 Report
Brief summary
The manuscript „Combined effects of submerged macrophytes and Daphnia Magna on bacterioplankton in eutrophic water” provides an insight into individual and combined effects of two different macrophytes and Daphnia Magna on the composition of planktonic bacteria evolution in eutrophic water. The well-prepared laboratory test with ten different scenarios lasted for 20 days and followed by a detailed analysis. According to the manuscript, laboratory experiments, that aim to reveal the combined effects of macrophytes and Daphnia, are very limited. Moreover, based on the available studies, the effectiveness of the proposed treatment is still questionable. From this point of view, the manuscript has high added values; however, in my opinion, it doesn’t fit well the main focus of the Sustainability journal. The performed research is related to topics such as sustainable use of resources or water pollution, but the manuscript doesn’t address these questions and linkages. The study would better fit to journals like Water or Ecologies (from MDPI). Other suitable journals might be Aquatic Botany or Inland Waters, for example.
In summary, the study represents an informative and detailed laboratory experiment that gives new insights about the effectiveness of Daphnia combined with submerged macrophytes to reduce algae development. We gain information as well about their interaction. In my opinion, the manuscript requires a major revision, which is related to how results, methods are presented. Furthermore, the discussion should be extended to link sustainability with the obtained conclusions.
Broad comments
The Introduction section gives a short summary of ecological methods to control algae abundance, such as through zooplanktons. The authors present recent studies and novel experiments; however, several statements are not well supported by references. I highlight these cases at the specific comments referring to line numbers.
Considering the Methods section, a detailed description is given by the manuscript. The laboratory experiment is sophisticated and accurate. The description of statistical analysis is somewhat superficial since, in most cases, the name of the software or program package is given instead of presenting the statistical tool. Furthermore, this section is too fragmented; namely, some sections are only 3-7 lines. I suggest to restructure this main section, and only three section levels should be used. For example, section 2.3.1 and the first part of 2.3.2.1 could be merged. Similarly, the second part of 2.3.2.1 and 2.3.2.2 can be condensed into one paragraph. I also suggest renaming the experiments to an easier traceability. For example: K – Control, DH – Daphnia 100 ind/L, DL – Daphnia 25 ind/L, C – Ceratophyllum, M – Myriophyllum. Following this, you will obtain for example: DHC – Daphnia 100 ind/L with Ceratophyllum, etc.
Regarding the Results section, in my opinion, the manuscript and the figures do not harmonize. A more detailed description and explanation would be useful for the figures to underpin the main messages. Furthermore, the figure captions are superficial, do not explain what we may observe in the figures. Just to mention one example, variable and units of axes are not given. For further details, please the specific comments.
The Discussion section synthesizes the main results. Firstly, I miss from this section is the earlier mentioned relationship with sustainability. Second, this was a truthful laboratory experiment; it would be great to read the authors’ thoughts about its applicability in practice. Pros and cons, strengths and weaknesses.
Specific comments and edits
L8: Repetition: “growth of”
L30: “biomanipulation is considered the most effective and ecological method to …” This a very strong statement; please rephrase or list references.
L32: The sentence: “An increase in plankton … of zooplankton” is strong and requires references.
L38: “many successful cases” – this requires references again.
L46: “In addition, other studies show…” Which studies? References are needed.
L52: “boast huge root systems” – What is the effect of root systems on bacterioplankton?
L54: Here, a third effect of macrophytes is given. This should also go into the listing.
L62: “This study aimed to evaluate the remediation …” I find this statement too strong. This study presents an important laboratory test. This is still far from remediation. Please, rephrase it!
L74: “Given the environmental value” – What do you mean by environmental value?
L87: “33 PVC” – What “33” does mean?
L91: “The initial water level was 20 L” – Liter is volume and not distance unit. What was the height and diameter of the barrel?
L92: “each index” Which indices? Water quality indicies from chapter 2.1?
L94: How was the barrel heated? Could stratification occur in the barrel? Was the sample mixed up in a daily manner?
L100: The distilled water had the same temperature, or was it colder? If yes, it could cause mixing.
L105: “three parallel test” – It is the first time mentioned that three parallel experimetns were conducted. This should be mentioned earlier.
L123: In this chapter, softwares are named instead of a description of the statistical procedures. References are also missing.
L131: Same as in the earlier chapter. Algorythms should be described very briefly.
L138: ANOVA and HSD. These are acronyms.
L139: “other statistical methods” – Namely?
L141: ‘a diagram was made” – What type and how?
L144: In this chapter, several indices are mentioned. These should be described in the methods sections.
L145: I believe you refer to Table 2.
L147: How these indices were calculated. At least, provide references.
L154: “Two commonly-used methods…” You name only one. Which is the second one?
L155: Figure 1 is referred, but nor explained, neither analyzed.
L172: Here, I believe you’re analyzing Figure 3. Refer to it.
L194: This first sentence belongs to Methods.
L196: How was water transparency determined?
L201: What is the correlation between WT and Chla?
L233: “provide major habitat” - In my opinion, this is one of the main result of the study. It would be great to read a further discussion about it.
L242: “assist recovery” – recovery from where to what?
L245-247: Were water quality parameters monitored during the experiment?
Figures and Tables
Figure 1 – What is plotted on the axis?
Figure 3 - What are the dots in the subfigures. Refer to subfigures in the caption. Chinese letters are shown in subfigure b).
Table 1: I suggest to rename the experiments as written above.
Table 2: The showed parameters should be described in the manuscript. Furthermore, analyses of section 3.1 refers to this figure. It would be worth considering the most important message of this table in a figure or graph.
Author Response
Response to reviewers’ comments:
(Reviewers comments are in Arial font and black color, our responses are in blue color)
Reviewers' comments:
The manuscript has been revised again carefully according to constructive comments and suggestions of the reviewers. We hope this revised manuscript can fit with the acceptable standard for Sustainability. We would like to express our hearted gratitude to the anonymous reviewers and the editor for their constructive comments and suggestions that really improved the manuscript greatly. We have replied all the comments from the reviewers into the revised version to improve the manuscript. Please see the following point-to-point answers with the marked-up manuscript version.
Reviewer1#
The Introduction section gives a short summary of ecological methods to control algae abundance, such as through zooplanktons. The authors present recent studies and novel experiments; however, several statements are not well supported by references. I highlight these cases at the specific comments referring to line numbers.
Considering the Methods section, a detailed description is given by the manuscript. The laboratory experiment is sophisticated and accurate. The description of statistical analysis is somewhat superficial since, in most cases, the name of the software or program package is given instead of presenting the statistical tool. Furthermore, this section is too fragmented; namely, some sections are only 3-7 lines. I suggest to restructure this main section, and only three section levels should be used. For example, section 2.3.1 and the first part of 2.3.2.1 could be merged. Similarly, the second part of 2.3.2.1 and 2.3.2.2 can be condensed into one paragraph. I also suggest renaming the experiments to an easier traceability. For example: K – Control, DH – Daphnia 100 ind/L, DL – Daphnia 25 ind/L, C – Ceratophyllum, M – Myriophyllum. Following this, you will obtain for example: DHC – Daphnia 100 ind/L with Ceratophyllum, etc.
Regarding the Results section, in my opinion, the manuscript and the figures do not harmonize. A more detailed description and explanation would be useful for the figures to underpin the main messages. Furthermore, the figure captions are superficial, do not explain what we may observe in the figures. Just to mention one example, variable and units of axes are not given. For further details, please the specific comments.
The Discussion section synthesizes the main results. Firstly, I miss from this section is the earlier mentioned relationship with sustainability. Second, this was a truthful laboratory experiment; it would be great to read the authors’ thoughts about its applicability in practice. Pros and cons, strengths and weaknesses.
Reply: Thanks for your constructive comments! Please see the point-to-point responses in the marked-up manuscript.
Specific comments and edits
L8: Repetition: “growth of”
Reply: Revised accordingly.
L30: “biomanipulation is considered the most effective and ecological method to …” This a very strong statement; please rephrase or list references.
Reply: We have revised the related descriptions and references. Thank you!
L32: The sentence: “An increase in plankton … of zooplankton” is strong and requires references.
Reply: References added accordingly.
L38: “many successful cases” – this requires references again.
Reply: References added accordingly.
L46: “In addition, other studies show…” Which studies? References are needed.
Reply: References added accordingly.
L52: “boast huge root systems” – What is the effect of root systems on bacterioplankton?
Reply: Boast huge root systems that can provide a living environment and attachment sites for the growth and reproduction of microorganisms, which would entail an increase in the diversity of microorganisms in water (Li et al. 2020). We have revised related descriptions accordingly. Thank you!
L54: Here, a third effect of macrophytes is given. This should also go into the listing.
Reply: Revised accordingly.
L62: “This study aimed to evaluate the remediation …” I find this statement too strong. This study presents an important laboratory test. This is still far from remediation. Please, rephrase it!
Reply: We have revised the sentences in line 113-115. Thank you!
L74: “Given the environmental value” – What do you mean by environmental value?
Reply: We have revised it in line 128. Thank you!
L87: “33 PVC” – What “33” does mean?
Reply: We have changed the related descriptions in line 153. Thank you!
L91: “The initial water level was 20 L” – Liter is volume and not distance unit. What was the height and diameter of the barrel?
Reply: The barrel is 30 cm diameter and a 30 cm height. We have revised it in line 157.
L92: “each index” Which indices? Water quality indices from chapter 2.1?
Reply: We have revised it in Line 158.
L94: How was the barrel heated? Could stratification occur in the barrel? Was the sample mixed up in a daily manner?
Reply: The heating rods is 20 cm height and the barrel is 30 cm height, and we slightly stirred the water in the barrel once every day between 8:00-9:00 a.m. to avoid the uneven heating. Thank you!
L100: The distilled water had the same temperature, or was it colder? If yes, it could cause mixing.
Reply: This study is carried out in June 2020, and the temperature change is around 20-28°C which is close to the experimental condition. In addition, we slightly stirred the water in the barrel once every day between 8:00-9:00 a.m. to avoid the uneven heating. Thank you!
L105: “three parallel test” – It is the first time mentioned that three parallel experiments were conducted. This should be mentioned earlier.
Reply: We have revised the related descriptions. Thank you!
L123: In this chapter, softwares are named instead of a description of the statistical procedures. References are also missing.
Reply: Thanks for the comments! We have added some software name in 2.4 Section.
L131: Same as in the earlier chapter. Algorythms should be described very briefly.
Reply: Revised accordingly.
L138: ANOVA and HSD. These are acronyms.
Reply: We have added the full name of the analysis method. Thank you!
L139: “other statistical methods” – Namely?
Reply: Redundancy analysis (RDA), we have revised the related descriptions. Thank you!
L141: ‘a diagram was made” – What type and how?
Reply: We have plotted RDA figure (Figure 4) by using the Canoco (Version 5.0) software. We have revised the descriptions to avoid confusions to the readers. Thank you!
L144: In this chapter, several indices are mentioned. These should be described in the methods sections.
Reply: We have added some related descriptions in Line 210-212. Thank you!
L145: I believe you refer to Table 2.
Reply: Revised accordingly.
L147: How these indices were calculated. At least, provide references.
Reply: Thanks for your suggestion, we have added some information where the calculation methods of these indices can be cited in line 212.
L154: “Two commonly-used methods…” You name only one. Which is the second one?
Reply: The second one is Non-MetricMulti-Dimensional Scaling (NMDS), and we have added in line 237. Thank you!
L155: Figure 1 is referred, but nor explained, neither analyzed.
Reply: We have added some related descriptions in line 241-246. Thank you!
L172: Here, I believe you’re analyzing Figure 3. Refer to it.
Reply: Revised accordingly.
L194: This first sentence belongs to Methods.
Reply: Revised accordingly.
L196: How was water transparency determined?
Reply: The Secchi disk depth (SDD) method was used to determine the transparency of water. Thank you!
L201: What is the correlation between WT and Chla?
Reply: In this figure, the bacterial community is the response variable, and the environmental factors such as WT and Chla are the explanatory variables. We did not do the correlation between environmental factors. Thank you!
L233: “provide major habitat” - In my opinion, this is one of the main result of the study. It would be great to read a further discussion about it.
Reply: Revised accordingly.
L242: “assist recovery” – recovery from where to what?
Reply: We have modified the related descriptions to avoid confusion. Thank you!
L245-247: Were water quality parameters monitored during the experiment?
Reply: We monitored the relevant data, but we did not put these data into this manuscript. We hope that will be fine. Thank you!
Figures and Tables
Figure 1 – What is plotted on the axis?
Reply: The axis represents the evolutionary distance among different microbials. We have added related information below Figure 1. Thank you!
Figure 3 - What are the dots in the subfigures. Refer to subfigures in the caption. Chinese letters are shown in subfigure b).
Reply: Asterisks indicates the significance among different microbials; *, p < 0.05; **, p < 0.01; ***, p < 0.001. Thank you!
Table 1: I suggest to rename the experiments as written above.
Reply: Revised accordingly.
Table 2: The showed parameters should be described in the manuscript. Furthermore, analyses of section 3.1 refers to this figure. It would be worth considering the most important message of this table in a figure or graph.
Reply: We have we have added some information where the calculation methods of these indices can be cited in line 211.
Thank you very much once again for your helpful comments!
Best Regards!
Bo Li (on behalf of all the co-authors)
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Associate Professor, Ph.D.
South China Agricultural University
College of Natural Resources and Environment
Department of Soil Science
483 Wushan Road, Tianhe District,
Guangzhou, 510642, China
MOB: +86 17612088475
Email: boli@scau.edu.cn
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Author Response File: 
Author Response.docx
Reviewer 2 Report
Bo Yang and colleges manuscript reported an experimental study which tested the responses of microbial communities to the addition of Daphnia magna and two macrophytes Ceratophyllum demersum and Myriophyllum spicatum in eutrophic water.
Major comments
The overall impression of the paper is that more effort is needed before it is ready to go.
The ecological relevance of the study and the experimental design are clear but in my eyes have poor support in the Results and Discussion section. Below the detailed comments are given.
Materials and Methods
The “OTUs taxonomic analysis” section has no references despite the qiime developers always ask to provide them and even give the example of Mat and Methods part on their site. Why the threshold of 70% is selected for taxonomy assignment? Seems a bit too low for studied ecosystem.
The “Statistical analysis” again has no references. Please, provide full information about the methods instead of “ and other statistical methods” in line 139.
Results
The Results section of papers based on in depth sequencing analysis of microbial communities usually provide the raw sequencing statistics at the beginning. Please, provide the information about the number of raw reads, merged pair-end reads, quality and chimera- filtered reads. Without that information there is no way to understand how well the whole experiment was done. Also the authors did not give the accession number of their reads in public databases which is a requirement for such kind of studies. Please, submit the sequenced reads to Genbank or EMBL dbs.
The Figures should provide the information about all presented symbols and abbreviations, f.e. there is no footnotes for asteriscs on Fig.2 and all hieroglyphs should be removed.
The data given in Table 2 is not in line with the description given in the text (line 149 and 151).
On barchart (which is actually the first figure and not Figure2) the authors provide the community composition at the phylum level then out of nowhere pick the top 20 species. Some names such as norank_f_Family1 or unclassified_k_norank are absolutely meaningless. Why are they left on Figures and discussed in the text? Such kind of negligence left the impression of draft manuscript but not the one you send to the journal. The table or the appropriate figure should be given with the whole taxonomy classification of selected species. If some of them have no taxonomic assignment at species level then left the name of higher taxonomic ranks.
Acidobacteria mentioned in the abstract as a result never appear in the further text.
I would recommend the authors to improve the mentioned problems, make the discussion part more consistent with the results, carefully read the text and fix the typo mistakes and then resubmit the paper again.
Author Response
(Reviewers comments are in Arial font and black color, our responses are in blue color)
Reviewers' comments:
The manuscript has been revised again carefully according to constructive comments and suggestions of the reviewers. We hope this revised manuscript can fit with the acceptable standard for Sustainability. We would like to express our hearted gratitude to the anonymous reviewers and the editor for their constructive comments and suggestions that really improved the manuscript greatly. We have replied all the comments from the reviewers into the revised version to improve the manuscript. Please see the following point-to-point answers with the marked-up manuscript version.
Reviewer2#
- Materials and Methods
The “OTUs taxonomic analysis” section has no references despite the qiime developers always ask to provide them and even give the example of Mat and Methods part on their site. Why the threshold of 70% is selected for taxonomy assignment? Seems a bit too low for studied ecosystem.
Reply: Thanks for your kind and suggestive comments. The threshold of taxonomy assignment is selected as 70%, which refers to the previous research. And the relevant references have been supplemented in the revised manuscript as you suggested in line 204-209.
The “Statistical analysis” again has no references. Please, provide full information about the methods instead of “ and other statistical methods” in line 139.
Reply: Thanks for your kind and suggestive comments. The methods and references have been supplemented in the revised manuscript as you suggested in line 220. Redundancy analysis (RDA) with a significance level of 0.05 so as to obtain the groups with significant.
Results
The Results section of papers based on in depth sequencing analysis of microbial communities usually provide the raw sequencing statistics at the beginning. Please, provide the information about the number of raw reads, merged pair-end reads, quality and chimera- filtered reads. Without that information there is no way to understand how well the whole experiment was done. Also the authors did not give the accession number of their reads in public databases which is a requirement for such kind of studies. Please, submit the sequenced reads to Genbank or EMBL dbs.
Reply: Thanks for your kind and suggestive comments. The raw sequenced reads have been submitted to Genbank in the revised manuscript as you suggested.
The Figures should provide the information about all presented symbols and abbreviations, f.e. there is no footnotes for asteriscs on Fig.2 and all hieroglyphs should be removed.
Reply: Thanks for your kind and suggestive comments. The notes of asteriscs on Fig.2 have been attached in the revised manuscript as you suggested.
The data given in Table 2 is not in line with the description given in the text (line 149 and 151).
Reply: Thanks for your kind and suggestive comments. The description of the high-density D. magna treatments (B) and the low-density D. magna treatments (A) were not consistent with the data given in Table 2. And these details have been corrected in the revised manuscript to make them consistent as you suggested.
On barchart (which is actually the first figure and not Figure2) the authors provide the community composition at the phylum level then out of nowhere pick the top 20 species. Some names such as norank_f_Family1 or unclassified_k_norank are absolutely meaningless. Why are they left on Figures and discussed in the text? Such kind of negligence left the impression of draft manuscript but not the one you send to the journal. The table or the appropriate figure should be given with the whole taxonomy classification of selected species. If some of them have no taxonomic assignment at species level then left the name of higher taxonomic ranks.
Reply: Thanks for your kind and suggestive comments. These names such as norank_f_Family1 or unclassified_k_norank have been deleted. And these details have been corrected in the revised manuscript as you suggested.
I would recommend the authors to improve the mentioned problems, make the discussion part more consistent with the results, carefully read the text and fix the typo mistakes and then resubmit the paper again.
Reply: Thanks for your kind and suggestive comments. And the other details have also been checked in the revised manuscript as you suggested.
Thank you very much once again for your helpful comments!
Best Regards!
Bo Li (on behalf of all the co-authors)
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Associate Professor, Ph.D.
South China Agricultural University
College of Natural Resources and Environment
Department of Soil Science
483 Wushan Road, Tianhe District,
Guangzhou, 510642, China
MOB: +86 17612088475
Email: boli@scau.edu.cn
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Author Response File: Author Response.docx
Reviewer 3 Report
I believe the manuscript is a timely and novel contribution to the field of Eutrophication control and should be considered for publication in Sustainability. It is an interesting paper; however, there are some lacking’s which I will try to explain below:
Specific comments:
- The title does not provide a targeted description of the content and should be changed.
- Language (grammar, wording, upper and lower case, typos, etc.) needs improvement by either a native speaker or a commercial service.
- Please be careful. Τhe word “D. magna” must be written in italics.
- Line 23: Please replace water with “water column” to better describe the process along the water as a function of the depth.
- Line 25: Not only in ponds, also in lakes. Please add “lakes” in the sentence.
- Line 26: …are in a stagnant state for a long time…? Please, you must rephrase your sentence. Ιn these closed ecosystems, the self-purification of water through the renewal of water bodies is negligible.
- Line 27: Eutrophication as a condition is not something neutral and it has negative effects. Therefore, the term “problem” better describes the phenomenon of eutrophication.
- Line 29: Please add a REF.
- Line 30: “…Among the scientific community…”. It is too general. Here you should briefly mention the other methods used to mitigate eutrophication.
- Line 39: Please add an appropriate reference.
- Line 48: Αlso here the reference is missing.
- Lines 67-69: What other physicochemical parameters were measured? Dissolved oxygen? pH? water transparency? etc. Please provide a table with these parameters.
- Did the water stirred during the whole experiment or not?
- Figure captions should be below the figure and not above it.
- Line 235: Environmental factors instead of environment factors
- Τhe relationship between environmental factors and bacterioplankton should be emphasized and clearly seen in the conclusions.
Author Response
(Reviewers comments are in Arial font and black color, our responses are in blue color)
Reviewers' comments:
The manuscript has been revised again carefully according to constructive comments and suggestions of the reviewers. We hope this revised manuscript can fit with the acceptable standard for Sustainability. We would like to express our hearted gratitude to the anonymous reviewers and the editor for their constructive comments and suggestions that really improved the manuscript greatly. We have replied all the comments from the reviewers into the revised version to improve the manuscript. Please see the following point-to-point answers with the marked-up manuscript version.
Reviewer3#
- The title does not provide a targeted description of the content and should be changed.
Reply: Thanks for your suggestion, we have modified the title with a targeted description accordingly.
- Language (grammar, wording, upper and lower case, typos, etc.) needs improvement by either a native speaker or a commercial service.
Reply: Thanks for your comments, and we have revised our language by a native speaker.
- Please be careful. Τhe word “D. magna” must be written in italics.
Reply: Revised accordingly.
- Line 23: Please replace water with “water column” to better describe the process along the water as a function of the depth.
Reply: Revised accordingly in line 34.
- Line 25: Not only in ponds, also in lakes. Please add “lakes” in the sentence.
Reply: Revised accordingly in line 37-38.
- Line 26: …are in a stagnant state for a long time…? Please, you must rephrase your sentence. Ιn these closed ecosystems, the self-purification of water through the renewal of water bodies is negligible.
Reply: Revised accordingly in line 37.
- Line 27: Eutrophication as a condition is not something neutral and it has negative effects. Therefore, the term “problem” better describes the phenomenon of eutrophication.
Reply: Revised accordingly in line 37.
- Line 29: Please add a REF.
Reply: Revised accordingly in line 40.
Val, H, & Smith. (2003). Eutrophication of freshwater and coastal marine ecosystems a global problem. Environmental Science & Pollution Research.
- Line 30: “…Among the scientific community…”. It is too general. Here you should briefly mention the other methods used to mitigate eutrophication.
Reply: Thanks for your comments! We have added some specific method for eutrophication mitigation in line 41-42.
- Line 39: Please add an appropriate reference.
Reply: Revised accordingly.
- Line 48: Αlso here the reference is missing.
Reply: Revised accordingly.
- Lines 67-69: What other physicochemical parameters were measured? Dissolved oxygen? pH? water transparency? etc. Please provide a table with these parameters.
Reply: Thanks for your comments, we have added some measured parameters in the 2.1 Section in line 120-123.
- Did the water stirred during the whole experiment or not?
Reply: Yes, the tested water has been stirred during the measuring process of some parameters.
- Figure captions should be below the figure and not above it.
Reply: Revised accordingly.
- Line 235: Environmental factors instead of environment factors
Reply: Revised accordingly in line 392.
- Τhe relationship between environmental factors and bacterioplankton should be emphasized and clearly seen in the conclusions.
Reply: Revised accordingly in line 421-423.
Thank you very much once again for your helpful comments!
Best Regards!
Bo Li (on behalf of all the co-authors)
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Associate Professor, Ph.D.
South China Agricultural University
College of Natural Resources and Environment
Department of Soil Science
483 Wushan Road, Tianhe District,
Guangzhou, 510642, China
MOB: +86 17612088475
Email: boli@scau.edu.cn
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Author Response File: Author Response.docx
Round 2
Reviewer 1 Report
Dear Authors,
Thank you for your point-by-point comments and for all the revisions you made. I have a couple of further concerns and suggestions to improve the manuscript.
The beginning of the new title, „D. magna with M. aquaticum combination” is grammatically incorrect. This is maybe better: „D. magna in combination with M. aquaticum…”
Heating and stirring information should be added to the manuscript, such as into the Collection Samples chapter.
In the first version, the description of statistical analysis was short-spoken. Instead of further details, the ms is completed by software names. Furthermore, some methods are given in the Results section instead of the Data analysis chapter.
The caption of Figure 2 is super long. Maybe, it would be better to add this description to the main text.
Finally, I still miss one or two paragraphs about the applicability of D. magna and macrophyte combinations in real circumstances to link the research to sustainability.
Thank you again for the revision and corrections!
Author Response
Response to reviewers’ comments:
(Reviewers comments are in Arial font and black color, our responses are in blue color)
Reviewers' comments:
The manuscript has been revised again carefully according to constructive comments and suggestions of the reviewers. We hope this revised manuscript can fit with the acceptable standard for Sustainability. We would like to express our hearted gratitude to the anonymous reviewers and the editor for their constructive comments and suggestions that really improved the manuscript greatly. We have replied all the comments from the reviewers into the revised version to improve the manuscript. Please see the following point-to-point answers with the marked-up manuscript version.
Reviewer1#
Thank you for your point-by-point comments and for all the revisions you made. I have a couple of further concerns and suggestions to improve the manuscript.
The beginning of the new title, „D. magna with M. aquaticum combination” is grammatically incorrect. This is maybe better: „D. magna in combination with M. aquaticum…”
Reply: Revised accordingly.
Heating and stirring information should be added to the manuscript, such as into the Collection Samples chapter.
Reply: Revised accordingly.
In the first version, the description of statistical analysis was short-spoken. Instead of further details, the ms is completed by software names. Furthermore, some methods are given in the Results section instead of the Data analysis chapter.
Reply: We have added the measurement methods for Chla and MC to improve the materials and methods section. In addition, due to the methods used in statistical analysis are currently commonly used internationally. Considering the brevity of the article, we did not do too many explanations.
The caption of Figure 2 is super long. Maybe, it would be better to add this description to the main text.
Reply: Revised accordingly. we move this paragraph to section 2.2.
Finally, I still miss one or two paragraphs about the applicability of D. magna and macrophyte combinations in real circumstances to link the research to sustainability.
Reply: we add a new section "4.5. application prospect" in our new manuscript.
Thank you again for the revision and corrections!
Thank you very much once again for your helpful comments!
Best Regards!
Bo Li (on behalf of all the co-authors)
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Associate Professor, Ph.D.
South China Agricultural University
College of Natural Resources and Environment
Department of Soil Science
483 Wushan Road, Tianhe District,
Guangzhou, 510642, China
MOB: +86 17612088475
Email: boli@scau.edu.cn
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Author Response File: Author Response.docx
Reviewer 2 Report
After first iteration some improvements was made by the authors. But I still believe that the quality of the article does not match the publication standard.
Major comments
1) As in my first review I am asking the authors to provide the raw sequencing statistics table. Again, without that information there is no way to understand how well the whole experiment was done. As an example see the following articles:
-Table 3 in Tonge DP, Pashley CH, Gant TW (2014) Amplicon –Based Metagenomic Analysis of Mixed Fungal Samples Using Proton Release Amplicon Sequencing. PLoS ONE 9(4): e93849. https://doi.org/10.1371/journal.pone.0093849
- Table 1 in Ivanova AA, Kulichevskaya IS, Merkel AY, Toshchakov SV and Dedysh SN (2016) High Diversity of Planctomycetes in Soils of Two Lichen-Dominated Sub-Arctic
Ecosystems of Northwestern Siberia. Front. Microbiol. 7:2065.
doi: 10.3389/fmicb.2016.02065
-Table S1 in Wegner CE, Liesack W. Microbial community dynamics during the early stages of plant polymer breakdown in paddy soil. Environ Microbiol. 2016 Sep;18(9):2825-42. doi: 10.1111/1462-2920.12815. Epub 2015 Apr 8. PMID: 25712035.
2) The authors reply to my comment regarding Genbank db was that the raw reads are now submitted. But where is the accession number in the revised manuscript?
3) The phylum Acidobacteria mentioned in the abstract as a result never appear in the main text. At least this information should be in the Results section
4) No_rank_f_Family1 still present on Fig 3A and Fig 4. Replace it with the higher rank meaningful taxon.
Author Response
(Reviewers comments are in Arial font and black color, our responses are in blue color)
Reviewers' comments:
The manuscript has been revised again carefully according to constructive comments and suggestions of the reviewers. We hope this revised manuscript can fit with the acceptable standard for Sustainability. We would like to express our hearted gratitude to the anonymous reviewers and the editor for their constructive comments and suggestions that really improved the manuscript greatly. We have replied all the comments from the reviewers into the revised version to improve the manuscript. Please see the following point-to-point answers with the marked-up manuscript version.
Reviewer2#
After first iteration some improvements was made by the authors. But I still believe that the quality of the article does not match the publication standard.
Major comments
1) As in my first review I am asking the authors to provide the raw sequencing statistics table. Again, without that information there is no way to understand how well the whole experiment was done. As an example see the following articles:
-Table 3 in Tonge DP, Pashley CH, Gant TW (2014) Amplicon –Based Metagenomic Analysis of Mixed Fungal Samples Using Proton Release Amplicon Sequencing. PLoS ONE 9(4): e93849. https://doi.org/10.1371/journal.pone.0093849
- Table 1 in Ivanova AA, Kulichevskaya IS, Merkel AY, Toshchakov SV and Dedysh SN (2016) High Diversity of Planctomycetes in Soils of Two Lichen-Dominated Sub-Arctic
Ecosystems of Northwestern Siberia. Front. Microbiol. 7:2065.
doi: 10.3389/fmicb.2016.02065
-Table S1 in Wegner CE, Liesack W. Microbial community dynamics during the early stages of plant polymer breakdown in paddy soil. Environ Microbiol. 2016 Sep;18(9):2825-42. doi: 10.1111/1462-2920.12815. Epub 2015 Apr 8. PMID: 25712035.
Reply: we add a new Table (Table S1) in our manuscript.
2) The authors reply to my comment regarding Genbank db was that the raw reads are now submitted. But where is the accession number in the revised manuscript?
Reply: It seems that we misunderstood your requirement in the last revision. We are so sorry that the raw reads can not be submitted due to the project requirements, and we hope that will be fine.
3) The phylum Acidobacteria mentioned in the abstract as a result never appear in the main text. At least this information should be in the Results section
Reply: we add a description about the Acidobacteria in section 4.1.
4) No_rank_f_Family1 still present on Fig 3A and Fig 4. Replace it with the higher rank meaningful taxon.
Reply: we remade the two Figures in our manuscript.
Thank you very much once again for your helpful comments!
Best Regards!
Bo Li (on behalf of all the co-authors)
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Associate Professor, Ph.D.
South China Agricultural University
College of Natural Resources and Environment
Department of Soil Science
483 Wushan Road, Tianhe District,
Guangzhou, 510642, China
MOB: +86 17612088475
Email: boli@scau.edu.cn
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Author Response File: Author Response.docx
Reviewer 3 Report
Thank you for sending the revised version and the author responses to my peer-review comments.
I was pleasantly surprised to see that the authors very carefully addressed the points raised that I had raised earlier.
Then dealt effectively with each point, corrected the items, and added valuable new content.
I recommend acceptance of the article now in its present form.
Author Response
Thanks for your kind support and wish all the best for you!
Best Regards!
Bo Li (on behalf of all the co-authors)
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Associate Professor, Ph.D.
South China Agricultural University
College of Natural Resources and Environment
Department of Soil Science
483 Wushan Road, Tianhe District,
Guangzhou, 510642, China
MOB: +86 17612088475
Email: boli@scau.edu.cn
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Round 3
Reviewer 2 Report
Dear authors,
I have no further comments regarding your article and thank you for all the revisions you have made.
Author Response
Thanks for your kind support and wish all the best for you!
Best Regards!
Bo Li (on behalf of all the co-authors)
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Associate Professor, Ph.D.
South China Agricultural University
College of Natural Resources and Environment
Department of Soil Science
483 Wushan Road, Tianhe District,
Guangzhou, 510642, China
MOB: +86 17612088475
Email: boli@scau.edu.cn
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
