Characteristics of Nitrogen Removal and Functional Gene Transcription of Heterotrophic Nitrification-Aerobic Denitrification Strain, Acinetobacter sp. JQ1004
Round 1
Reviewer 1 Report
Comments and Suggestions for Authors
Journal: Water
Manuscript ID: water-3032303
Title: Characteristics of Nitrogen Removal and Functional Genes Transcription of Heterotrophic Nitrification-Aerobic Denitrification Strain, Acinetobacter sp. JQ1004
To Authors:
In this study, the authors simulated the biomass proliferation pattern of heterotrophic nitrification and aerobic denitrification (HNAD) Acinetobacter sp. JQ1004 isolated in the previous work by the Logistic model. In addition the expression of functional genes was investigated to characterize the cellular molecular and potential degradation mechanism. In fact, the nitrogen removal by HNAD strain JQ1004 has been widely investigated in the literature. However, the kinetics and mechanism analysis of the process have not been reported in detailed. Therefore, I believe the manuscript can be considered for publication after a major revision. There are some flaws and errors (e.g. language problems, models evaluation etc) which should be address before consideration for publication. Detailed comments are listed below:
1. A concern that needs to be addressed is the need for grammatical changes and errors that are beyond the scope of the reviewer. This includes the absence of articles of speech (examples: a, an, the), which is a common occurrence among non-native English speakers. The authors also mix past and present tense in their paragraphs. It may be helpful to have someone else review the text to catch these mistakes.
ABSTRACT:
2. The novelty and main objectives of the work should be clearly mentioned in the abstract.
INTRODUCTION:
3. Please add one paragraph (before your last paragraph) and explain briefly about the N removal mechanism by HNAD and highlight the important of modeling of the process. These are your novelty (I guess) and you should focus on them.
MATERIALS ANS METHODS:
4. Explain about model fitting process and how did you defined the fitting parameters?
RESULTS AND DISCUSSION:
5. Again you should clearly discuss about model evaluation. What was the models error (MRE, R2, etc)
6. Provide Experimental data vs. Model prediction data (with R2)
7. The quality of Fatigues should be improved.
Comments on the Quality of English Language
A concern that needs to be addressed is the need for grammatical changes and errors that are beyond the scope of the reviewer. This includes the absence of articles of speech (examples: a, an, the), which is a common occurrence among non-native English speakers. The authors also mix past and present tense in their paragraphs. It may be helpful to have someone else review the text to catch these mistakes.
Author Response
To reviewer #1:
We sincerely thank you for giving us the opportunity to revise and resubmit our manuscript. We feel sorry that we do have some inadequacies in our previous manuscript. Your thoughtful comments have contributed a lot to improve the quality of our manuscript. As you are concerned, there are several problems that need to be addressed. According to your suggestions, we have made extensive corrections to our previous draft. All changes/additions to the manuscript are given in blue text. The detailed corrections are listed below.
Specific comments from reviewer #1:
Comment 1: A concern that needs to be addressed is the need for grammatical changes and errors that are beyond the scope of the reviewer. This includes the absence of articles of speech (examples: a, an, the), which is a common occurrence among non-native English speakers. The authors also mix past and present tense in their paragraphs. It may be helpful to have someone else review the text to catch these mistakes.
Response: Thank you for your suggestion. We have checked and corrected the grammar and spelling errors in our manuscript. The changes/additions are given in blue text. We hope it could be up to your standard this time.
Comment 2: The novelty and main objectives of the work should be clearly mentioned in the abstract.
Response: Thanks for your comments. According to your suggestions, we have revised the abstract to highlight the novelty and main objectives of this work. The changes/additions are given in blue text. We hope it could be up to your standard this time.
Comment 3: Please add one paragraph (before your last paragraph) and explain briefly about the N removal mechanism by HNAD and highlight the important of modeling of the process. These are your novelty (I guess) and you should focus on them.
Response: Thanks for your comments. According to your suggestions, we have re-discussed and explained about the mechanism of nitrogen removal by HNAD and highlighted the important of modeling of the process in the last paragraph of the section 3.4. The changes/additions are given in blue text. We hope it could be up to your standard this time.
Comment 4: Explain about model fitting process and how did you defined the fitting parameters?
Response: Thank you for your comments. Bacterial suspension was inoculated into the liquid HM or DM to conduct the experiments on the degradation of different nitrogen by strain JQ1004. With the culture time (t, h) as the horizontal coordinate and N concentration (mg/L) as the vertical coordinate, the degradation law of different nitrogen by HN-AD bacteria was plotted (Experimental data). The degradation kinetics model on different N was fitted using a modified Compertz model (Model prediction data).
Comment 5: Again you should clearly discuss about model evaluation. What was the models error (MRE, R2, etc)
Response: Thank you for your comments. The R2 of the fitted curves for the degradation of ammonia and nitrate were 0.997 and 0.985, respectively. As for degrading ammonia, the model errors were 0.47 and 0.35 for Rm and t0, respectively. As for nitrate, the model errors were 0.32 and 1.13 for Rm and t0, respectively. We have re-discussed the model evaluation, and the changes/additions are given in blue text. We hope it could be up to your standard this time.
Comment 6: Provide Experimental data vs. Model prediction data (with R2)
Response: Thank you for your valuable comments. According to your suggestions, we have made the change in Fig. 1 by adding and labelling the Experimental data vs. Model prediction data. We hope it could be up to your standard this time.
Comment 7: The quality of Fatigues should be improved.
Response: Are you referring to the quality of figures? Thank you for your reminding. We have improved the quality of figures and converted it to a new format for re-uploading. We hope the revised figures could be up to your standard this time.
Author Response File: 
Author Response.pdf
Reviewer 2 Report
Comments and Suggestions for Authors
Substantial revision is required, rest is okay
Comments for author File: Comments.pdf
Author Response
To reviewer #2:
Thank you so much for your professional review work on our manuscript. Your thoughtful comments and constructive suggestions have contributed a lot to improve the quality of our manuscript. According to your suggestion, we have made some modifications to our previous draft. We tried our best to revise the “Introduction” section and highlighted the significance of our findings. And the Keywords were revised based on your suggestions. Besides, we have checked and corrected the grammar and spelling errors of the manuscript. All changes/additions to the manuscript are given in blue text. We hope the revised manuscript could be up to your standard this time.
Specific comments from reviewer #2:
Question 1: What is the main question addressed by the research?
Comment: The research gap and objective are not defined clearly. Author should mention the main objective and aim of the study in the last para in the Introduction part.
Response: Thank you for your comments. According to your suggestions, we have revised the section of the “Introduction” to elucidate the main objective and aim of this work. All changes/additions to the manuscript are given in blue text. We hope the revised manuscript could be up to your standard this time.
Question 2: What specific improvements should the authors consider regarding the methodology? What further controls should be considered?
Comment : • Authors should be thorough revised and elaborate the entire methodology.
- The author should also well write the procedure of the isolation technique.
- In section 2.2 Nitrogen balance analysis the author should give full methodology with references.
- In section 2.4 why the author uses sodium succinate and ammonium sulfate as carbon and nitrogen sources give an explanation and also add references.
- In line 106- what is OD600, also explain it??
- On line 119 what is SYBR green data 1 and also describe 2⸹⸹CT methods give full detail.
Response: Thank you for your careful check. According to your suggestion, we have revised the section of ‘2. Materials and Methods’. In section 2.4, the sodium succinate and ammonium sulfate have been used as carbon and nitrogen sources respectively because of their simple molecular structure and easy bioavailability. Besides, OD600 refers to the absorption value of a solution at a wavelength of 600 nm. One of its important applications is to use the absorption of bacteria to measure the concentration of bacterial culture medium, so as to estimate the growth of bacteria, so it is usually used to refer to the density of bacterial cells. The method of SYBR Green Ι refers to the application of a non-specific DNA-binding dye with fluorescence to detect the accumulated amplification products during PCR. In the PCR reaction system, excessive SYBR fluorescent dye is added. After SYBR fluorescent dye is specifically incorporated into the DNA double strand, the fluorescence signal is emitted, while the SYBR dye molecule in the unincorporated chain does not emit any fluorescence signal, so as to ensure that the increase of fluorescence signal is completely synchronized with the increase of PCR product. The 2-△△CT method refers to an algorithm that calculates the relative expression of the target gene after fluorescence quantitative PCR. All changes/additions to the manuscript are given in blue text. We hope the revised manuscript could be up to your standard this time.
Question 3: Are the references appropriate?
Comment : Not yet, add some references to justify your findings and also site some recent
articles. Also may follow the pattern and flow of the presentation??
Response: Thank you for your comments. According to your suggestions, we have revised the section of ‘References’. Some recent articles were sited to support our findings. All changes/additions to the manuscript are given in blue text. We hope the revised manuscript could be up to your standard this time.
Question 4: additional comments on the tables and figures and quality of the data.
Comment: • In line 178:- Add equation number.
- In line 241- Correct figure (a).
- For Figure-1. The author should provide the complete data.
- Please mention more recent references in the discussion section compared with the present study.
Response: Thanks for your careful check. We have accurately added the equation number to the cited position in the manuscript. The figures and tables have been modified as your suggestions. Some recent references were added to compare with the present study. All changes/additions to the manuscript are given in blue text. We hope the revised manuscript could be up to your standard this time.
Author Response File: Author Response.pdf
Round 2
Reviewer 1 Report
Comments and Suggestions for Authors
The manuscript has bee revised appropriately and can be published by Water.
