C058 and Other Functional Microorganisms Promote the Synthesis of Extracellular Polymer Substances in Mycelium Biofloc
Round 1
Reviewer 1 Report
The authors fail to provide proper framework for some of the experiments performed, see for instance lines 58/59 “It can be seen from Fig. 1 that the EPS content of the experimental reactor was higher than that of the control reactor under various HRTs”…what was the intended goal of the experiment? From the materials and methods it is not clear what was the medium in the control and in the experimental reactor…one only understand the difference when reading later that in the experimental reactor CO58 flocs were used whereas the control inoculated the Pearl River water that contained rich species of bacteria. Which bacteria? How were they chosen? Please clarify!
Overall, the authors are suggested to organize the text of the manuscript so that the experimental set-up designed is understandable and how it can contribute to achieve the intended goals
Line 40 what is the meaning of acronyms TN and TP, they are not given in the text
What is meant by “the law of PN and PS contents in EPS were the same as that of EPS” in particular what is meant by “the law”?
The authors state that “HRT was adjusted by changing the flow speed, which produced different shear forces. The appropriated shear force was benefit for the EPS secretion in aerobic granular sludge, but excessive shear force was not conducive to the sludge stability”. Please use flow rate rather than flow speed. Can it be assumed that increased the flow rate decreases HRT and increases shear force? The results obtained when control and experimental reactor are considered are contradictory. In the latter, EPS is at a minimum for the lowest HRT whereas in the former EPS is at a maximum for the lowest HRT. Maybe other factors should be considered
Error bars or similar should be added to the figures to clarify the statistical significance of the data
Line 73 what are “first and second stages”? Unclear
What is meant by SVI? The acronym was never defined throughout the text
It is strongly recommended the assistance of a native English speaker, there are too many glitches that compromise the understanding of the matters addressed
Two examples
There is an error in the title “Mechanism of EPS synthesis in C058 mycilium biofloc” should be “Mechanism of EPS synthesis in C058 mycelium biofloc”
Please mind the tense in the introduction lines 26 to 35, present tense should be used
Author Response
Question 1: The authors fail to provide proper framework for some of the experiments performed, see for instance lines 58/59 “It can be seen from Fig. 1 that the EPS content of the experimental reactor was higher than that of the control reactor under various HRTs”…what was the intended goal of the experiment? From the materials and methods it is not clear what was the medium in the control and in the experimental reactor…one only understand the difference when reading later that in the experimental reactor CO58 flocs were used whereas the control inoculated the Pearl River water that contained rich species of bacteria. Which bacteria? How were they chosen? Please clarify!
Question 2: Overall, the authors are suggested to organize the text of the manuscript so that the experimental set-up designed is understandable and how it can contribute to achieve the intended goals.
Reply: The intended goal of this study is to explore whether the addition of C058 promotes the synthesis of EPS. Hence, comparing with an ordinary biofloc (OBF, without C058), this study analyzed the changes of extracellular polymer substances content of the mycelium biofloc (MBF, with C058). And then the EPS in MBF and CBF are compared to explore the expected goal of this study. Some key points for the experimental design are supplemented in the 3.2.2 section belonged to the Materials and Methods.
Question 3: Line 40 what is the meaning of acronyms TN and TP, they are not given in the text.
Reply: Now the full names are used instead of its abbreviations.
Question 4: What is meant by “the law of PN and PS contents in EPS were the same as that of EPS” in particular what is meant by “the law”?
Question 5: The authors state that “HRT was adjusted by changing the flow speed, which produced different shear forces. The appropriated shear force was benefit for the EPS secretion in aerobic granular sludge, but excessive shear force was not conducive to the sludge stability”. Please use flow rate rather than flow speed. Can it be assumed that increased the flow rate decreases HRT and increases shear force? The results obtained when control and experimental reactor are considered are contradictory. In the latter, EPS is at a minimum for the lowest HRT whereas in the former EPS is at a maximum for the lowest HRT. Maybe other factors should be considered.
Question 6: Error bars or similar should be added to the figures to clarify the statistical significance of the data.
Question 7: Line 73 what are “first and second stages”? Unclear
Question 8: What is meant by SVI? The acronym was never defined throughout the text.
Question 9: It is strongly recommended the assistance of a native English speaker, there are too many glitches that compromise the understanding of the matters addressed.
Two examples
There is an error in the title “Mechanism of EPS synthesis in C058 mycilium biofloc” should be “Mechanism of EPS synthesis in C058 mycelium biofloc”.
Please mind the tense in the introduction lines 26 to 35, present tense should be used.
Reviewer 2 Report
The article "Mechanism of EPS synthesis in C058 mycilium biofloc" presents good results, but the following suggestions can improve the quality of the article, to be accepted and published in this journal.
Line 40 and 41 in recent years?
Why were 36, 18, and 11 hour times used?
In Figure 1 (and in the other figures on the bar) it is necessary to add the standard deviation of the measurements to also verify the differences between the statistics and the values presented.
Other characterizations would be differentiated to be added as zeta potentials, and other technical techniques to enrich the article.
In SEM the images are in different magnifications (10 or 30 um).
Figure 7 can be replaced by standard tables.
Author Response
Review Report 2
The article "Mechanism of EPS synthesis in C058 mycilium biofloc" presents good results, but the following suggestions can improve the quality of the article, to be accepted and published in this journal.
Question 1: Line 40 and 41 in recent years?
Reply: Although bioaugmentation method have been extensively studied and applied in activated sludge technology, its usage in biofloc technology is just in recent years as indicated by literature [12,13,14], since the biofloc technology was just developed by an Israeli scientist named Avnimelech in 1999 based on the activated sludge technology.
Question 2: Why were 36, 18, and 11 hour times used?
Reply: The initial HRT 36h was designed according to the growth period of C058 strain, and HRT 18h was half of that, but HRT 11h (not 9h) was used for a reason of the limitation of the flow rate range (40-210ml/min) of the peristaltic pump (model: KCPPRO2-N16) that used in this experience.
Question 3: In Figure 1 (and in the other figures on the bar) it is necessary to add the standard deviation of the measurements to also verify the differences between the statistics and the values presented.
Reply: Error bars are added in Figure 1 and Figure 2.
Question 4: Other characterizations would be differentiated to be added as zeta potentials, and other technical techniques to enrich the article.
Reply: In this study, the main contents are EPS contents, floc microstructure and microflora composition, so scanning electron microscope and high-throughput sequencing are used. Other characterizations such as Zeta potential are deficient in the test. If necessary, this point will be taken into account in the subsequent study. Thank you for your kind suggestion.
Question 5: In SEM the images are in different magnifications (10 or 30 um).
Reply: Now the SEM images are in the same magnification of 30 um.
Question 6: Figure 7 can be replaced by standard tables.
Reply: The stacked bar chart in Figure 7 is more intuitive than a standard table, and also it keeps a consistent form with Figure 5. Here Figure 7 is retained. Thank you for your kind suggestion.
Reviewer 3 Report
The following report is based on my review of the manuscript entitled “Mechanism of EPS synthesis in C058 mycilium biofloc”, with manuscript number catalysts-1715614. The manuscript fits within the scope of Catalysts and interesting. However, the following shortcomings have been pointed out and need to be addressed properly for further improvement of the manuscript. They are:
1. It is suggested to modify the title “Mechanism of EPS synthesis in C058 mycilium biofloc”. The word “modelling”. The EPS synthesis mechanism type should be captured in the title.
2. Kindly remove all abbreviations from the Abstract and Keyword
3. In the introduction, it is suggested to merge paragraphs 1 and 2. This is because paragraph 2 seems to be too short.
4. The novelty and practical applicability of this study should be highlighted more in the introduction section as essential research components such as existing problem, methodology adopted and expected outcomes are not clear in the article. The sequence of the introduction section is also not appropriate. It should be improved. Author(s) should refer to these articles highlighted below and several others relating to EPS synthesis from reputable journals to enrich and enhance the introduction section of the manuscript. Hence, they should be cited appropriately. These are:
· https://doi.org/10.1007/s13399-022-02431-2
· https://doi.org/10.1016/j.jwpe.2021.102178
5. Authors tried to present the structure of the article at the end of the introduction. However, it is not well structured. Kindly improve on that. The novelty of the work alongside the objectives should be highlighted more in the last paragraph of the introduction section. Presently, it is too shallow.
6. The “Materials and method” section should come before “Results and discussion of result”.
7. The conclusion section should either be in a single paragraph or main findings of the study be highlighted in bullet forms.
8. It is weird that author did not present a picture of the mycelium biofloc (MBF) bioaugmented by Cordyceps strain C058.
Finally, the manuscript should be modified according to above said comments and be thoroughly reviewed again before accepting it for publication.
Author Response
The following report is based on my review of the manuscript entitled “Mechanism of EPS synthesis in C058 mycilium biofloc”, with manuscript number catalysts-1715614. The manuscript fits within the scope of Catalysts and interesting. However, the following shortcomings have been pointed out and need to be addressed properly for further improvement of the manuscript. They are:
Question 1: It is suggested to modify the title “Mechanism of EPS synthesis in C058 mycilium biofloc”. The EPS synthesis mechanism type should be captured in the title.
Reply: Now the title “C058 and other functional microorganisms promote the synthesis of extracellular polymer substances in mycelium biofloc” may cover the EPS synthesis mechanism.
Question 2: Kindly remove all abbreviations from the Abstract and Keyword.
Reply: All abbreviations are removed from the Abstract and Keyword.
Question 3: In the introduction, it is suggested to merge paragraphs 1 and 2. This is because paragraph 2 seems to be too short.
Reply: Yes, paragraphs 1 and 2 can be merged.
Question 4: The novelty and practical applicability of this study should be highlighted more in the introduction section as essential research components such as existing problem, methodology adopted and expected outcomes are not clear in the article. The sequence of the introduction section is also not appropriate. It should be improved. Author(s) should refer to these articles highlighted below and several others relating to EPS synthesis from reputable journals to enrich and enhance the introduction section of the manuscript. Hence, they should be cited appropriately. These are:
https://doi.org/10.1007/s13399-022-02431-2
https://doi.org/10.1016/j.jwpe.2021.102178
Reply: Now the existing problem and the intended goal of this study, in particular the practical applicability of the present study, is highlighted more in the introduction section.
Several articles (literature 5, 11, and 21) are cited to enrich and enhance the introduction section.
Question 5: Authors tried to present the structure of the article at the end of the introduction. However, it is not well structured. Kindly improve on that. The novelty of the work alongside the objectives should be highlighted more in the last paragraph of the introduction section. Presently, it is too shallow.
Reply: Thank you for your kind suggestion. As stated in the introduction, the present study aims at illustrating the water purification effect of MBF through EPS synthesis, which may be helpful for its practical applicability. Hence, the introduction section is not a long description. It is worth mentioning the suage of bioaugmentation method in biofloc technology is just in recent years, and the biofloc technology as well as its existing problems have been highlighted in the introduction section. Additionally, the literature 5, 11, and 21 are cited to optimize the introduction structure.
Question 6: The “Materials and method” section should come before “Results and discussion of result”.
Reply: According to the format of this journal, "Materials and method " section should be placed after " Results and discussion of result ".
Question 7: The conclusion section should either be in a single paragraph or main findings of the study be highlighted in bullet forms.
Reply: Now the conclusion section are in bullet forms.
Question 8: It is weird that author did not present a picture of the mycelium biofloc (MBF) bioaugmented by Cordyceps strain C058.
Reply: Fig.4 a, c and e are the pictures of the mycelium biofloc bioaugmented by C058 under various hydraulic retention times.
The manuscript has been modified according to above comments. And thanks very much for your kind suggestions.
Round 2
Reviewer 1 Report
The authors managed to address the main criticisms to the original version
Reviewer 3 Report
Authors have made significant improvement on the manuscript. All queries have been addressed.
