The Effect of the Artificial Reef on the Structure and Function of Sediment Bacterial Community
, Guobao Chen 1,2,3,4,5, Xue Feng 1,2,3,4,5, Yan Liu 1,2,3,4,5 and Pimao Chen 1,2,3,4,5Round 1
Reviewer 1 Report
This article dealt with prokaryotic community structure and function in the artificial reef area. This study investigated the influence of artificial reef, on prokaryotic plankton in the sediment. It is useful to re-cognition artificial reef along the coastline. I think this study was well organized, while it needs minor revision before it is accepted for publishing.
I think there is no need to use the specific statistical methods in the abstract, such as Redundancy analysis (RDA), FAPROTAX results, just directly use statistical results showed that….Detailed comments are listed below.
1. Line 46, the unit “empty cubic meters” was confusion. Please check that all units for the manuscript.
2. The “artificial reefs” has been already mentioned and defined its abbreviation in the introduction, and it can be used the abbreviation subsequently.
3. When reporting stats results, if subsequent statistical results you report are from the same test, you should state the name in the first instance.
4. The water depth was misuse as “deep” in the figure 6 and 8. Please check that word for the manuscript.
5. Please check the capital or small letters all the paper, such as line 255 “Spearman Correlation coefficient”.
6. In Figure 8, please add a title to your color scale, to explain what is being shown.
7. The legend of figure 8, for environment factor Change to for environmental factor.
8. The font should be unified in the graph, please check the full text font in the figure of this paper.
9. In these figures of this paper, the genus name should keep in italics. Please check the generic name in the text and the figure. Such as Fig. 6.
10. Please add missing spaces before the parentheses.
11. Pay attention to the cohesion of sentences in the text. Some closely connected sentences are segmented. Such as Line 420.
12. There omission the URL of the software in the thirty-fifth reference.
13. In the references, there have several defects, such as the 38th reference, here the enter key seems unnecessarily.
14. Revise the reference format according to the requirements of the “Instructions for Authors” carefully. Specifically, in Line 238, 258, there is “Error! Reference source not found.). Based”, please recheck it.
Author Response
On behalf of my co-authors, we thank you very much for giving us an opportunity to revise our manuscript, and we also appreciate reviewers very much for their positive and constructive comments and suggestions on our manuscript.
Point 1: I think there is no need to use the specific statistical methods in the abstract, such as Redundancy analysis (RDA), FAPROTAX results, just directly use statistical results showed that. Detailed comments are listed below.
Response 1: We have deleted the specific statistical methods and directly reported statistical results in the abstract. Thanks for your advise.
Point 2: Line 46, the unit “empty cubic meters” was confusion. Please check that all units for the manuscript.
Response 2: We have revised the unit “empty cubic meters” to “cubic meters” and inspected the units throughout. Thanks for your advise.
Point 3: The “artificial reefs” has been already mentioned and defined its abbreviation in the introduction, and it can be used the abbreviation subsequently.
Response 3: We have revised the words and inspected the abbreviation throughout. Thanks for your advise.
Point 4: When reporting stats results, if subsequent statistical results you report are from the same test, you should state the name in the first instance.
Response 4: We have revised the description when report the statistical results throughout. Thanks for your advise.
Point 5: The water depth was misuse as “deep” in the figure 6 and 8. Please check that word for the manuscript.
Response 5: We have revised the spelling throughout. Thanks for your advise.
Point 6: Please check the capital or small letters all the paper, such as line 255 “Spearman Correlation coefficient”.
Response 6: We have revised the spelling throughout. Thanks for your advise.
Point 7: In Figure 8, please add a title to your color scale, to explain what is being shown.
Response 7: We have added a title to the color scale. Thanks for your advise.
Point 8: The legend of figure 8, for environment factor Change to for environmental factor.
Response 8: We have revised the spelling throughout.. Thanks for your advise.
Point 9: The font should be unified in the graph, please check the full text font in the figure of this paper.
Response 9: We have revised the font throughout.. Thanks for your advise.
Point 10: In these figures of this paper, the genus name should keep in italics. Please check the generic name in the text and the figure. Such as Fig. 6.
Response 10: We have revised the font throughout.. Thanks for your advise.
Point 11: Please add missing spaces before the parentheses.
Response 11: We have added missing spaces before the parentheses follow the adivse.
Point 12: Pay attention to the cohesion of sentences in the text. Some closely connected sentences are segmented. Such as Line 420.
Response 12: We have deleted the enter key and conneted the two sentence. Thanks for your advise.
Point 13: In the references, there have several defects, such as the 38th reference, here the enter key seems unnecessarily.
Response 13: We have revised the invalid format follow the adivse.
Point 14: There omission the URL of the software in the thirty-fifth reference.
Response 14: We have revised the added the missing URL follow the adivse.
Point 15: Revise the reference format according to the requirements of the “Instructions for Authors” carefully. Specifically, in Line 238, 258, there is “Error! Reference source not found.). Based”, please recheck it.
Response 15: We have revised error follow the adivse.
Author Response File: 
Author Response.docx
Reviewer 2 Report
I strongly recommend this paper for publication, albeit I have a few comments and a few things need improvement. Here is a list of them:
Line 122 – instead of total genome DNA should be total genomic DNA
Line 138 which NovaSeq was used? 6000 or Novaseq X plus? I assume that it was NovaSeq 6000 since 250 bp reads were obtained. However, in line 188 authors mention Illumina MiSeq sequencing. So my question is: which platform was used?
Line 169 record PRJNA880993 is not available, but I hope that before publication the data will be made available?
Figure 3, on x-axis should be Shannon, not shanno
Line 223, 230, 238, 258 Error! Reference source not found
Line 269 Woeseia should be written in italic
Author Response
On behalf of my co-authors, we thank you very much for giving us an opportunity to revise our manuscript, and we also appreciate reviewers very much for their positive and constructive comments and suggestions on our manuscript.
Point 1: Line 122 – instead of total genome DNA should be total genomic DNA.
Response 1: We have revised the “genome” to “genomic”. Thanks for your advise.
Point 2: Line 138 which NovaSeq was used? 6000 or Novaseq X plus? I assume that it was NovaSeq 6000 since 250 bp reads were obtained. However, in line 188 authors mention Illumina MiSeq sequencing. So my question is: which platform was used?
Response 2: We used the NovaSeq 6000 platform in the present study, and the “Illumina MiSeq” was
a misstatement. We have revised the error. Thanks for your advise.
Point 3: Line 169 record PRJNA880993 is not available, but I hope that before publication the data will be made available?
Response 3: We will change the release of th data date before publication. Thanks for your advise.
Point 4: Figure 3, on x-axis should be Shannon, not shanno
Response 4: We have revised the spelling mistake in the Figures. Thanks for your advise.
Point 5: Line 223, 230, 238, 258 Error! Reference source not found
Response 5: We have revised the errors. Thanks for your advise.
Author Response File: Author Response.docx
Reviewer 3 Report
The authors have brought out a highly informative piece of work relating to the artificial reefs and sediment bacteria. The following suggestions may be looked into:
The authors infer that the present study and the associated parameters could throw light on the trophic transfer of energy in marine ranching food webs. Also, there are mentions of pathogenic bacteria present in the sediment system. Kindly clarify whether these could negatively impact the biota of the reef ecosystem. Also, does the study provide any insights into the microbial loop system and regenerated production? Please clarify the envisaged connectivity to fishery resources as well.
Discussion section: Line 318 -The statement is confusing as to whether the author is referring to the current study or the previously reported study. Please clarify.
Line 340 - The statement is too abrupt. Please establish connectivity to the study and modify the statement accordingly.
Author Response
On behalf of my co-authors, we thank you very much for giving us an opportunity to revise our manuscript, and we also appreciate reviewers very much for their positive and constructive comments and suggestions on our manuscript.
Point 1: In this paper, the key issues were theoretically reviewed and well-structured. However, there are several typographical, spelling, and punctuation errors. These should be corrected throughout the text. or The article needs to be carefully proofread.
Response 1: We thank the referee for spotting these typographical, spelling, and punctuation errors, both of which are now corrected in the revised manuscript.
Point 2: Abstract is well structured but need further improvement with aim and scope.
Response 2: We have improved the aim and scope in the abstract. Thanks for your advise.
Point 3: Add some details on marine ranching and why it is important to construct. Add the impact of the discharge of urban and industrial wastewater and overfishing.
Response 3: We have add some details about the wastewater and overfishing in the introduction. Thanks for your advise.
Point 4: The research methods followed in the research paper should be addressed with proper references.
Response 4: Some references are added in corresponding positions for the methods part. Thanks for your advise.
Author Response File: Author Response.docx
Reviewer 4 Report
The authors compared the dynamics of the bacterial composition and potential biogeochemical functions of a ten-year artificial reef, a one-year new artificial reef, and buck seawater. This project provides insights into the influence of artificial reefs on the associated microbiome and marine environment. However, the current manuscript is insufficient to publish in this journal for the following reasons. The author used the general microbiome pipeline to analyze the data without some optimization. The legends of the figures were not detailed enough, and the data analysis method was not given in detail. The flow of the manuscript should be more fluent. The discussion section should discuss the combined results rather than explain each result section.
Comments:
1. How does the author explain that the differences in microbiomes and parameters are not due to geographic differences but somewhat artificial reefs?
2. According to Figure 1 and chapter 2.1, “Samples were collected at 250–300 m intervals along the center of each area”. Is it too far from the artificial reef? Why not collect the mud samples just near the artificial reefs?
3. Lines 59-60, explain the influence of calcium hydroxide ions, iron, and silicate released into the marine environment.
4. The authors should provide more details about how artificial reefs affect biogeochemical and nutrient cycling processes.
5. Could the authors explain why they did not measure calcium hydroxide ions, iron, and silicate, since the concrete may cause those elements to be released into the seawater?
6. Suggestion: it’s better to test the phytoplankton community as well in the future. Phytoplankton is the primary productivity of the marine ecosystem.
7. The authors did not mention any tools or R packages for MNDS, RDA, and network plots.
8. Chapter 2.5: Did the authors remove the primers and barcodes when processing the raw data? It did not mention.
9. The authors should change the database for taxonomy annotation. GreenGene 13.8 is no longer being maintained since 2013. To get more accurate and convincing results, please use SILVA 138, updated in 2021.
10. Line 196, please explain the most abundant ASVs and rare abundant ASVs.
11. Line 198, the 7.63% present numbers or relative abundance?
12. How many reads did the authors get after QC control? Could the authors provide rarefaction plots to show the sequencing depth is sufficient?
13. The authors need a barplot to show the relative abundance of each sample.
14. Figure 3, y-axis “Shanno*n”
15. Lines 202-207, the authors explained Fig3b first. The way to present results should follow the order of figures.
16. Line 223, reference error
17. Fig4, it is a mess of showing all the taxonomy levels for LefSe. The authors did not show the cladogram figure even though they mentioned it in the legend.
18. The authors have three different groups for analyzing. I would like to suggest using ternary plots for visualization. It is in the vegan package in R.
19. Figure 5, explain the ellipse. Could the authors explain why they chose the NMDS (non-metric algorithm) rather than PcoA (metric algorithm)?
20. Line 238, reference error
21. Lines 245-251, include “(Figure 6)”.
22. Line 250, N*O*3
23. The authors could try MENA (Molecular ecological network analyze)http://129.15.40.240/mena/(Ye Deng et al., 2012 BMC Bioinformatics) to build highly co-occurring bacterial taxa (modules) and compute the Correlation between module eigenvalue and environmental parameter.
24. Line 338 sediments’, Line 339 samples’
25. Lines 379-392, confused the audience when the authors described all the taxonomy levels together. For example, the authors do not need to mention both Moraxellaceae and Acinetobacter since Acinetobacter belongs to Moraxellaceae. The authors need a better flow to present data.
Author Response
On behalf of my co-authors, we thank you very much for giving us an opportunity to revise our manuscript, and we also appreciate reviewers very much for their positive and constructive comments and suggestions on our manuscript. we have optimized the legend, figures, and structure of the manuscript. We wish the revised manuscript would like to meet the publication requirements of this journal.
Point 1: How does the author explain that the differences in microbiomes and parameters are not due to geographic differences but somewhat artificial reefs?
Response 1: Thanks for your question. Prior to the construction of the artificial reef area at this place, we have already carried out a background investigation and selected the area of artificial reef construction, there was no significant difference in the background characteristics including environmental parameters and microbial characteristics between the regions.
Point 2: According to Figure 1 and chapter 2.1, “Samples were collected at 250–300 m intervals along the center of each area”. Is it too far from the artificial reef? Why not collect the mud samples just near the artificial reefs?
Response 2: The sampling sites were all located in artificial reef areas, with an area of about 500 × 500m, and thousands of artificial reefs were thrown in the marine ranching sea area. samples were near the artificial reefs. We have supplemented this part in the method section. Thanks for your advise.
Point 3: Lines 59-60, explain the influence of calcium hydroxide ions, iron, and silicate released into the marine environment.
Response 3: The calcium hydroxide ions may influence the pH of the enviroment, and the CaO and CaCO₃ particles were found to have significant effects on marine species (Brooks et al., 2020). Iron deficiency limits phytoplankton growth in many high-latitude and equatorial regions of the world ocean (Zhang et al., 2019). Iron is found in two oxidation states within the hydrosphere: reduced ferrous iron (Fe II) and oxidized ferric iron (Fe III). Soluble ferrous iron, which is toxic, rapidly oxidizes to ferric iron under oxygenated conditions (Han et al., 2021). Silicate could act as an inducer of laccase production by Acinetobacter sp . and effect the growth of the marine diatom(Anusaraporn et al., 2022; Hemalatha et al., 2012). Thanks for your question.
Point 4: The authors should provide more details about how artificial reefs affect biogeochemical and nutrient cycling processes.
Response 4: Tons of concrete reefs on the sea floor changed the flow field of the sea floor. The complex flow field near ARS not only provides a suitable activity area for various marine organisms but also promotes the exchange of various nutrients and enhances the ecological effect of ARS. The transport and exchange of sediment nutrients can be promoted by upwelling. Thanks for your advise.
Point 5: Could the authors explain why they did not measure calcium hydroxide ions, iron, and silicate, since the concrete may cause those elements to be released into the seawater?
Response 5: With the extension of reef time and the damage of reef, the main structural concrete and reinforcement of reef may release these elements under the corrosion of seawater. However, if the quality of concrete meets the requirements of construction standards and the reinforcement is wrapped inside the concrete, thus, the iron, silicate and calcium oxide released by artificial reef will be relatively low in the short term. On the other hand, this present study focused on the interaction of artificial reefs with harmful heavy metals and nutrients, therefore we did not extensively measure these elements. Thanks for the question.
Point 6: Suggestion: it’s better to test the phytoplankton community as well in the future. Phytoplankton is the primary productivity of the marine ecosystem.
Response 6: We agree with the reviewer that the phytoplankton community was important for the primary productivity of the marine ecosystem. We will report the phytoplankton in the future. Thanks for the reviewer’s suggestion.
Point 7: The authors did not mention any tools or R packages for MNDS, RDA, and network plots.
Response 7: We used the R (version 3.3.1) vegan packages for the MNDS, RDA, and igraph packages for network plots. These information have been supplemented in the method. Thanks for the reviewer’s advise.
Point 8: Chapter 2.5: Did the authors remove the primers and barcodes when processing the raw data? It did not mention.
Response 8: We have trimmed to the V4 region bound by the 515FmodF/806RmodR primer pair. We have described it in the menthod. Thanks for the reviewer’s advise.
Point 9: The authors should change the database for taxonomy annotation. GreenGene 13.8 is no longer being maintained since 2013. To get more accurate and convincing results, please use SILVA 138, updated in 2021.
Response 9: It is relatively accurate due to manual sorting. Many researchers still choose to use the database (Kaczmarek et al., 2020; Procter et al., 2022). The commonly used families, genera and species of the seven phyla are used for classification, which is easy to understand and read. It is also the default species annotation database of QIIME software. PICRUST analysis tool, that is, the analysis of predicting microbial community functions based on 16S rRNA high-throughput sequencing results, is also developed based on Greengne database. The downside is false positives will be higher for the SILVA. We would love to used the SILVA in the future. Thanks for the reviewer’s advise.
Point 10: Line 196, please explain the most abundant ASVs and rare abundant ASVs.
Response 10: The most abundant ASVs refers to the largest number of feature sequence and the rarest refers to the least number of feature sequence. We have revised the sentence make it more clear. Thanks for the reviewer’s advise.
Point 11: Line 198, the 7.63% present numbers or relative abundance?
Response 11: The 7.63% refers to ASVs numbers. Thanks for the reviewer’s advise.
Point 12: How many reads did the authors get after QC control? Could the authors provide rarefaction plots to show the sequencing depth is sufficient?
Response 12: We identified a total of 650,977 high-quality reads across all 12 samples, ranging from 48,128 to 62,576 reads per sample. These reads were clustered into 11,517 ASVs. The rarefaction curve plots was shown as a supplement to show the sequencing depth is sufficient. Thanks for the reviewer’s advise.
Point 13: The authors need a barplot to show the relative abundance of each sample.
Response 13: The barplot of the relative abundance of each sample was shown as a supplement to follow the adivse.
Point 14: Figure 3, y-axis “Shanno*n”
Response 14: We have revised the error to the figure. Thanks for the reviewer’s advise.
Point 15: Lines 202-207, the authors explained Fig3b first. The way to present results should follow the order of figures.
Response 15: We have revised the order when report the figure. Thanks for the reviewer’s advise.
Point 16: Line 223, reference error
Response 16: We have revised the error. Thanks for the reviewer’s advise.
Point 1: Fig4, it is a mess of showing all the taxonomy levels for LefSe. The authors did not show the cladogram figure even though they mentioned it in the legend.
Response 17: The figure showed the taxonomy of genus level for LefSe. However, the second generation sequencing can only obtain the taxonomy information at the level families or orders for some species. We have modified the legend. Thanks for the reviewer’s advise.
Point 18: The authors have three different groups for analyzing. I would like to suggest using ternary plots for visualization. It is in the vegan package in R.
Response 18: The ternary of the three groups was shown in the 3.2 section. Thanks for the reviewer’s advise.
Point 19: Figure 5, explain the ellipse. Could the authors explain why they chose the NMDS (non-metric algorithm) rather than PcoA (metric algorithm)?
Response 19: The ellipse refers to a 95% confidence interval. PCoA and NMDS are used to reflect the sample distance matrix relationship. The difference is that NMDS focuses more on reflecting the ranking relationship of values in the distance matrix, and weakens the absolute difference of values. PCoA is based on the distance matrix calculated from species abundance. NMDS is a nonlinear model, which can overcome some shortcomings of linear models and better reflect the nonlinear structure of ecological data. When there are too many samples or species, NMDS will be more accurate. Non metric multidimensional calibration method statistics is a sort method suitable for ecological research.
Point 20: Line 238, reference error
Response 20: We have revised the error in the manuscript. Thanks for the reviewer’s advise.
Point 21: Lines 245-251, include “(Figure 6)”.
Response 21: The Figure S3 was a supplement to show the environmental parameters in clusters and correlate them with the sediment microbial communities. If the reader would love to more intuitive to find the which environmental factors have significant effects on special genera or phyla of microorganisms. The heatmap plot make the data more intuitive and contrasting. Thanks for the reviewer’s advise.
Point 22: Line 250, N*O*3
Response 22: We have revised the error in the manuscript. Thanks for the reviewer’s advise.
Point 23: The authors could try MENA (Molecular ecological network analyze) http://129.15.40.240/mena/ (Ye Deng et al., 2012 BMC Bioinformatics) to build highly co-occurring bacterial taxa (modules) and compute the Correlation between module eigenvalue and environmental parameter.
Response 23: Due to the MENA currently can not handle dataset with less than 8 samples. This present study was 4 samples each group, so the MENA could not deal with the interaction network within the group. We would love to try the analyze tool in the future. Thanks for the reviewer’s advise.
Point 24: Line 338 sediments’, Line 339 samples’
Response 24: We have revised the error in the manuscript. Thanks for the reviewer’s advise.
Point 25: Lines 379-392, confused the audience when the authors described all the taxonomy levels together. For example, the authors do not need to mention both Moraxellaceae and Acinetobacter since Acinetobacter belongs to Moraxellaceae. The authors need a better flow to present data.
Response 25: This LEfSe analysis was based on the taxonomy of genus level. However, the second generation sequencing can only obtain the taxonomy information at the level families or orders for some species. The Moraxellaceae screened in this LEfSe analysis is actually a different genus from Acinetobacter, but the available data only identify him at the family level. So we can only infer his features by describing Moraxellaceae. This expression is also used treatment in other literature (Zhou et al., 2022).We have modified the describe make it more distinct. Thanks for the reviewer’s advise.
References
Anusaraporn, S., Dolphen, R., & Thiravetyan, P. (2022). Importance of laccase enzyme and triiodide for gold leaching from silicate ore by marine bacterium Acinetobacter sp. Process Safety and Environmental Protection, 161, 788–800. https://doi.org/10.1016/j.psep.2022.03.054
Brooks, S. J., Georgantzopoulou, A., Johansen, J. T., & Mengede, M. (2020). Determining the risk of calcium oxide (CaO) particle exposure to marine organisms. Marine Environmental Research, 156, 104917. https://doi.org/10.1016/j.marenvres.2020.104917
Han, C., Kim, H.-J., Lee, J.-S., Sakakura, Y., & Hagiwara, A. (2021). Species-specific effects of iron on temperate and tropical marine rotifers in reproduction, lipid and ROS metabolisms. Chemosphere, 277, 130317. https://doi.org/10.1016/j.chemosphere.2021.130317
Hemalatha, A., Karthikeyan, P., Manimaran, K., Anantharaman, P., & Sampathkumar, P. (2012). Effect of Temperature on the Growth of Marine Diatom, Chaetoceros simplex (Ostenfeld, 1901) with Different Nitrate: Silicate Concentrations. Asian Pacific Journal of Tropical Biomedicine, 2(3, Supplement), S1817–S1821. https://doi.org/10.1016/S2221-1691(12)60501-2
Kaczmarek, Ł., Roszkowska, M., Poprawa, I., Janelt, K., Kmita, H., Gawlak, M., Fiałkowska, E., & Mioduchowska, M. (2020). Integrative description of bisexual Paramacrobiotus experimentalis sp. Nov. (Macrobiotidae) from republic of Madagascar (Africa) with microbiome analysis. Molecular Phylogenetics and Evolution, 145, 106730. https://doi.org/10.1016/j.ympev.2019.106730
Procter, M., Kundu, B., Sudalaimuthuasari, N., AlMaskari, R. S., Saeed, E. E., Hazzouri, K. M., & Amiri, K. M. A. (2022). Microbiome of Citrullus colocynthis (L.) Schrad. Reveals a Potential Association with Non-Photosynthetic Cyanobacteria. Microorganisms, 10(10), Article 10. https://doi.org/10.3390/microorganisms10102083
Zhang, R., Kelly, R. L., Kauffman, K. M., Reid, A. K., Lauderdale, J. M., Follows, M. J., & John, S. G. (2019). Growth of marine Vibrio in oligotrophic environments is not stimulated by the addition of inorganic iron. Earth and Planetary Science Letters, 516, 148–155. https://doi.org/10.1016/j.epsl.2019.04.002
Zhou, Z., Meng, H., Gu, W., Li, J., Deng, M., & Gu, J.-D. (2022). High-throughput sequencing reveals the main drivers of niche-differentiation of bacterial community in the surface sediments of the northern South China sea. Marine Environmental Research, 178, 105641. https://doi.org/10.1016/j.marenvres.2022.105641
Author Response File: Author Response.docx
Reviewer 5 Report
The manuscript was carefully reviewed. In this study, the authors investigated the effect of the artificial reef on the structure and function of the sediment bacterial community. The authors have presented an interesting paper. The work is well presented, but many issues need to be solved before publishing. Some of my comments on this paper are shown below:
In this paper, the key issues were theoretically reviewed and well-structured. However, there are several typographical, spelling, and punctuation errors. These should be corrected throughout the text. or The article needs to be carefully proofread.
Abstract is well structured but need further improvement with aim and scope.
Add some details on marine ranching and why it is important to construct. Add the impact of the discharge of urban and industrial wastewater and overfishing.
The research methods followed in the research paper should be addressed with proper references.
Please add the significance of the research in the conclusion part.
Author Response
On behalf of my co-authors, we thank you very much for giving us an opportunity to revise our manuscript, and we also appreciate reviewers very much for their positive and constructive comments and suggestions on our manuscript.
Point 1: The authors infer that the present study and the associated parameters could throw light on the trophic transfer of energy in marine ranching food webs. Also, there are mentions of pathogenic bacteria present in the sediment system. Kindly clarify whether these could negatively impact the biota of the reef ecosystem. Also, does the study provide any insights into the microbial loop system and regenerated production? Please clarify the envisaged connectivity to fishery resources as well.
Response 1: Some pathogenic bacteria present are opportunistic pathogens in the sediment system. They may be lead to biota of the reef ecosystemreefs organisms disease in a specific environment conditions. For example, when the proliferating organisms exceed the environmental capacity or the local water environment suffers from the discharge of sewage and so on, the marine biota may be more susceptible to the infection of pathogenic bacteria. Marine sediments have an important impact on fisheries resources. For example, if too much organic matter is concentrated in the sediments, poor flow exchange could lead to rapid oxygen consumption in the sea area, which in turn could lead to oxygen deprivation in the bottom seawater, or produce toxic compounds such as hydrogen sulphide, which are detrimental to fisheries, and microbes in the sediments, which are also affected by the reef environment, affect the recycling of these organic matter and sulphides, therefore, the reef sediments have a significant impact on the fishery resources in the Marine Rangeing area.
Thanks for your advise.
Point 2: Discussion section: Line 318 -The statement is confusing as to whether the author is referring to the current study or the previously reported study. Please clarify.
Response 2: The satement refers to previously reported study. We have revised the sentence. Thanks for your advise.
Point 3: Line 340 - The statement is too abrupt. Please establish connectivity to the study and modify the statement Accordingly
Response 3: We have rewritten the section of the sentence to enhance the coherence of sentences. Thanks for your advise.
Author Response File: Author Response.docx
Round 2
Reviewer 4 Report
This manuscript is much improved. Authors have done all the changes according to my comments. Good job.
