Comparative Analysis of Metagenomic (Amplicon and Shotgun) DNA Sequencing to Characterize Microbial Communities in Household On-Site Wastewater Treatment Systems

Round 1

Reviewer 1 Report

1) There are many studies using both amplicon sequencing and shotgun sequencing, thus, the authors should make an effort for up-to-date literature to show the gaps in knowledge and the significance of this study.

2) What are the influents of OWTS in this study? Real wastewater vs. synthetic wastewater?

3) TKN, NH3, TSS, and COD were mentioned in the method but did not present and relate to the microbial community in the results.

4) There is a lack of interpretation for the result of the microbial community in section 3.2

5) There are many methods for analyzing microbial communities using shotgun sequencing. Here, the authors only applied Diamond-Blastx against the NCBI database for analysis. Please explain why? It is probably the reason for the higher richness of the microbial community. There are other methods such as Kaiju, mOTUs2, etc. for microbial characterization.

6) A core microbial community for OWTS should be presented in the result.

 

7) The conclusion is very lengthy, please make it concise and clear. 

Author Response

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Author Response File: Author Response.docx

Reviewer 2 Report

The concept of the study is basic and the draft has written according to the journal's format, however few fuzzy statements are quoted in this article and seems appropriate references are not given. The specific comments are appended here for correction. I recommend the manuscript is not suitable for publication process

1) Did the author perform any culturing process.

2) how did the author confirm the strain…… not appropriate explanation

3)  What is the basic theme of identification of strain and fails to insist the treatment procedure.

4) Just the author uptake the sample and predict the sequencing procedure … is there any further usage of strain in further treatment process.

5) The author must improve the discussion part.

6) should rewrite the conclusion part…

 

Author Response

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Author Response File: Author Response.docx

Reviewer 3 Report

This manuscript describes “metagenomics analysis of household onsite wastewater treatment systems using 16S and shotgun sequencing”.

 

1.    In introduction, in addition add more information regarding shotgun and 16s sequencing with advantages and disadvantages.

2.    In this manuscript, microbial analysis from different treatment trains was done. However, there is no information on the title. If possible, revise the title, which provides more information to match the content of the manuscript.

3.    Add more information why metagenomics analysis is crucial in onsite wastewater treatment systems in introduction and conclusion, even proper place.

4.    Compare data of species level between shotguns and 16 S sequencing if the data is available.

Author Response

see attached

Author Response File: Author Response.docx

Round 2

Reviewer 2 Report

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