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We agree with the reviewer’s comments and make Figure S4 as Figure 4 in this revised version.
| 2 | 1 |
I would suggest to avoid citing supplementary figures at the beginning of the chapter. See for example chapter 2.4. Given that 2.4 chapter discusses data of supplementary figure 4 I would suggest to make figure S4 as Figure 4 of the manuscript.
| 1 | 2 |
toxins14060409_makarova
| 1 |
We marked the inoculation sites with red arrows in Figure 1.
| 2 | 1 |
To futher clarify the infection methods a supplementary figure detailing graphically the modes of inoculation with figures would make the paper extremely useful for the community
| 1 | 2 |
toxins14060409_makarova
| 1 |
We added more details about the strain and cited the following paper. (Pag.10 Line 402-403) Zhang, Y.; Li, A.; Zhu, S.; Li, L.; He, X.; Sun, Z.; Li, T. Basal Rachis Internode Injection (BRII): A novel inoculation method to evaluate wheat resistance to Fusarium head blight. Phytopathology, 2021, 111, 1670-1674, doi:10.1094/PHYTO-11-20-0488-R. Point 4: Mycotoxin data analysis should be provided (I guess is policy of mdpi to make raw data available together with the publication) Response 4: Thanks!
| 2 | 1 |
Identity of the strain used for infection should be confirmed: a multilocus species characterisation is needed to confirm the species of the strain or a reference to a publication where the strain was described and appropriately characterised.
| 1 | 2 |
toxins14060409_makarova
| 1 |
Thanks! We uploaded the raw mycotoxin data as suggested.
| 2 | 1 |
Mycotoxin data analysis should be provided (I guess is policy of mdpi to make raw data available together with the publication)
| 1 | 2 |
toxins14060409_makarova
| 1 |
Following the suggestion of this reviewer, we have removed the movie in the revised version.
| 2 | 1 |
The largest concern is regarding the colocalization of IFITM3 and Rab11A. It seems (as the authors state) that these represent abortive events (Figure 7). Therefore the inclusion of the movie examining the colocalization of Rab11A GFP, IFITM3-SNAP, and labeled virion seems completely unnecessary. In addition, the movie does not provide any details on cellular features and a viewer is left wondering where the foci are coming from and going. This reviewer suggests removing the movie or alternatively including labels on the movie of cellular features and including a scale bar.
| 1 | 2 |
v11060548_makarova
| 1 |
We show indirect immunofluorescence analysis with anti-IFITM3 and anti-NP antibodies of HSAEpCs in Figure S6. No unspecific signal is detected in the cytosol of HSAEpCs, indicating that the observed cytoplasmic NP stain is specific. Nuclear NP stain is detected at 10 h p.i., and this may indeed indicate delayed replication compared to A549 cells as suggested by the reviewer, but no direct side-by-side comparison of replication kinetics in different cells was performed.
| 2 | 1 |
b. The NP stain in HSAEpCs is only shown to be cytoplasmic, inclusion of an uninfected control would demonstrate specificity of the staining. At later timepoints is the NP stain nuclear as expected, is this simply a delay in replication?
| 1 | 2 |
v11060548_makarova
| 1 |
The IFITM3 images were acquired in the cytosol distant from the nucleus and absent from the plasma membrane. We included this information in the respective figure legend of Figure 3 and 7. Figure 3: “STED images (raw data) show representative subcellular regions located in the cytosolic part distant from the nucleus and absent from the plasma membrane as used for cluster analysis.” Figure 7: “Images show regions located in the cytosolic part distant from the nucleus and absent from the plasma membrane.”
| 2 | 1 |
c. Labels or zoomed out images from those presented in Figure 3 and 7 are needed to orient the reader to where within a cell the colocalization is observed.
| 1 | 2 |
v11060548_makarova
| 1 |
We now include the Western Blot analysis suggested by the reviewer as Figure S2B and thank the reviewer for suggesting this important control.
| 2 | 1 |
d. Figure 2B - Western blot examining levels of IFITM3 should include 24 hpi to correlate with the data presented in Figure 1.
| 1 | 2 |
v11060548_makarova
| 1 |
We have made the suggested changes.
| 2 | 1 |
Figure 1 should be labeled as to what treatments are used in panels A-E. As it is now, the figure on its own cannot possibly be interpreted without the legend.
| 1 | 2 |
v11060548_makarova
| 1 |
We now include an immunoblot showing IFITM3 levels in whole-cell lysates from A549 wildtype and IFITM3 knock-down cells as Figure S1D.
| 2 | 1 |
Figure 2A: The siRNA control should be shown. qPCR is acceptable, but an IFITM3 Western would be better, particularly since the antibody is readily available.
| 1 | 2 |
v11060548_makarova
| 1 |
To address the reviewer’s concern, we performed statistical analysis of three independent experiments shown in Figure 2B, lower panel. Statistical analysis of mean values of the determined protein amount for IFITM3 shows no statistically significant differences (e.g. p-value of 6 h pi compared to control is 0.5224). We included this information in the figure legend of Figure 2.
| 2 | 1 |
Figure 2B: These data need to be improved. The IFITM3 blot shows wide variability with no pattern from one hour to the next, and yet the conclusion is that there is no change in IFITM3 levels during this time period.
| 1 | 2 |
v11060548_makarova
| 1 |
If this is not acceptable we will make the changes in final revision. We have not made this change as we show that the anti-NP antibody is highly specific and shows negligible signal in the uninfected controls in Figure 1A. A representative example of non-infected HSAEpC is shown in the supplement.
| 2 | 1 |
Figure 4: Uninfected control should be shown in the main text.
| 1 | 2 |
v11060548_makarova
| 1 |
We have made the requested changes.
| 2 | 1 |
Figure 4: Label the figure so that it can be interpreted independent of the legend.
| 1 | 2 |
v11060548_makarova
| 1 |
We have made the requested changes to figure 5A/C and Figure 7.
| 2 | 1 |
Figure 5A/C: Label the figure so that it can be interpreted independent of the legend.
| 1 | 2 |
v11060548_makarova
| 1 |
We used a fluorescently tagged Rab11 because the anti-Rab11 antibodies were not sufficient to label recycling endosomes. However, IFITM3 is detected at native levels and this compromise allowed us to evaluate its localization. Overexpressed IFITM3 was only used for the live cell microscopy shown in the movie, which has now been removed (see reply to reviewer 1).
| 2 | 1 |
Figure 7: I am not sure that this figure is a useful addition to the paper as it suffers from exactly the problems that the authors claim to overcome in their study, i.e., the use of overexpressed IFITM3 and an overexpressed Rab11 marker protein, both of which may show unnatural localization due to the overexpression.
| 1 | 2 |
v11060548_makarova
| 1 |
Based on the reviewer´s suggestion, we have now included again the reference showing the infection increase of IAV in A549 IFITM3 knockdown cells and have changed the wording to make the statement clearer. “The mechanism by whichIFITM3 impedes IAV infection was divergent in previous studies and inhibition was mostly described upon IFITM3 over-expression [37, 38, 51]. It was demonstrated that IFITM3 knockdown in A549 cells increases infection rate [37].” Since this paper is primarily studying IFITM localization and clustering at endosomes/lysosomes it should reference studies in which endocytic localization motifs of the IFITMs were identified and characterized (Jia, Cell Microbiol, 2014; Chesarino, JBC, 2015; Li, JBC, 2015)
| 2 | 1 |
Lines 227/228: This sentence is misleading as the references cited are not contradictory, nor are they unclear as to IFITM3 possessing antiviral activity. Furthermore, the authors need to examine the literature more carefully, as previous studies have indeed shown that IFITM3 knockdown in A549 cells increases influenza virus infection, e.g., Lin, Cell Reports, 2013.
| 1 | 2 |
v11060548_makarova
| 1 |
All the references have been added and we now include this aspect of the IFITM3 mechanism in the discussion of the paper. We thank the reviewer for the comment.
| 2 | 1 |
Since this paper is primarily studying IFITM localization and clustering at endosomes/lysosomes it should reference studies in which endocytic localization motifs of the IFITMs were identified and characterized (Jia, Cell Microbiol, 2014; Chesarino, JBC, 2014; Li, JBC, 2015) Line 65 states that subsequent studies have failed to confirm an association between SNP rs12252 and severe influenza.
| 1 | 2 |
v11060548_makarova
| 1 |
Lines 61 ff. were changed pointing out the relevance of SNP rs12252-C and SNP rs34481144-A. References have been added.
| 2 | 1 |
Line 65 states that subsequent studies have failed to confirm an association between SNP rs12252 and severe influenza. This may be true, but far more studies have confirmed this link than have refuted it. Further, the studies which failed to find an association were generally performed in populations in which the SNP is almost non-existent. Additionally, a second SNP in the IFITM3 promoter has also been linked to severe flu. For a review of these studies, see Zani, Current Clin Microbiol Reports, 2018, though the primary articles should be cited.
| 1 | 2 |
v11060548_makarova
| 1 |
We thank the reviewer for pointing out the inaccuracy in the references. The statement "...IFITM3 elevates the level of cholesterol on late endosomes and lysosomes thereby restricting early IAV infection" is based on the publication of Kühnl et al. 2018 (mBio). The reference was replaced. We have also complemented the introductory part with the aspect of fusion inhibition through the amphipathic helix of IFITM3.
| 2 | 1 |
Lines 67-73 The inhibition of virus membrane fusion by IFITM3 has been attributed to the presence of a palmitoylated amphipathic helix within IFITM3 (Chesarino, EMBO Reports, 2017). This helix and neighboring palmitoylation sites are among the most highly conserved residues among IFITMs from all species. Additionally, the role of cholesterol in IFITM3’s mechanism of action has been largely disproven by the field, and reference 52 seems to have been misused in line 73.
| 1 | 2 |
v11060548_makarova
| 1 |
The supplemental figures only include experiments that reproduce previously published work or are not absolutely necessary to support the major and most novel findings of the paper. We therefore suggest to keep these data in the supplement.
| 2 | 1 |
Supplemental figures should be included in the main text.
| 1 | 2 |
v11060548_perova
| 1 |
We show indirect immunofluorescence analysis with anti-IFITM3 and anti-NP antibodies of HSAEpCs in Figure S6. No unspecific signal is detected in the cytosol of HSAEpCs, indicating that the observed cytoplasmic NP stain is specific. Nuclear NP stain is detected at 10 h p.i., and this may indeed indicate delayed replication compared to A549 cells as suggested by the reviewer, but no direct side-by-side comparison of replication kinetics in different cells was performed.
| 2 | 1 |
The NP stain in HSAEpCs is only shown to be cytoplasmic, inclusion of an uninfected control would demonstrate specificity of the staining. At later timepoints is the NP stain nuclear as expected, is this simply a delay in replication?
| 1 | 2 |
v11060548_perova
| 1 |
The IFITM3 images were acquired in the cytosol distant from the nucleus and absent from the plasma membrane. We included this information in the respective figure legend of Figure 3 and 7. Figure 3: “STED images (raw data) show representative subcellular regions located in the cytosolic part distant from the nucleus and absent from the plasma membrane as used for cluster analysis.” Figure 7: “Images show regions located in the cytosolic part distant from the nucleus and absent from the plasma membrane.” d.
| 2 | 1 |
Labels or zoomed out images from those presented in Figure 3 and 7 are needed to orient the reader to where within a cell the colocalization is observed.
| 1 | 2 |
v11060548_perova
| 1 |
We now include the Western Blot analysis suggested by the reviewer as Figure S2B and thank the reviewer for suggesting this important control.
| 2 | 1 |
Figure 2B - Western blot examining levels of IFITM3 should include 24 hpi to correlate with the data presented in Figure 1.
| 1 | 2 |
v11060548_perova
| 1 |
We have made the suggested changes.
| 2 | 1 |
Figure 1 should be labeled as to what treatments are used in panels A-E. As it is now, the figure on its own cannot possibly be interpreted without the legend.
| 1 | 2 |
v11060548_perova
| 1 |
We now include an immunoblot showing IFITM3 levels in whole-cell lysates from A549 wildtype and IFITM3 knock-down cells as Figure S1D.
| 2 | 1 |
Figure 2A: The siRNA control should be shown. qPCR is acceptable, but an IFITM3 Western would be better, particularly since the antibody is readily available.
| 1 | 2 |
v11060548_perova
| 1 |
To address the reviewer’s concern, we performed statistical analysis of three independent experiments shown in Figure 2B, lower panel. Statistical analysis of mean values of the determined protein amount for IFITM3 shows no statistically significant differences (e.g. p-value of 6 h pi compared to control is 0.5224). We included this information in the figure legend of Figure 2.
| 2 | 1 |
Figure 2B: These data need to be improved. The IFITM3 blot shows wide variability with no pattern from one hour to the next, and yet the conclusion is that there is no change in IFITM3 levels during this time period.
| 1 | 2 |
v11060548_perova
| 1 |
Figure 2B shows that the amount of IFITM3 is not significantly altered up to 6 h p.i. IFITM3 protein increased significantly after longer infection times such as 24 h p.i. (Fig. S2B), as was previously described by others.
| 2 | 1 |
Figure 3A: Why is there so much IFITM3 present in the uninfected cells? This does not appear to increase post infection as was shown previously.
| 1 | 2 |
v11060548_perova
| 1 |
If this is not acceptable we will make the changes in final revision. We have not made this change as we show that the anti-NP antibody is highly specific and shows negligible signal in the uninfected controls in Figure 1A. A representative example of non-infected HSAEpC is shown in the supplement.
| 2 | 1 |
Figure 4: Uninfected control should be shown in the main text.
| 1 | 2 |
v11060548_perova
| 1 |
We have made the requested changes.
| 2 | 1 |
Figure 4: Label the figure so that it can be interpreted independent of the legend.
| 1 | 2 |
v11060548_perova
| 1 |
We have made the requested changes to figure 5A/C and Figure 7.
| 2 | 1 |
Figure 5A/C: Label the figure so that it can be interpreted independent of the legend.
| 1 | 2 |
v11060548_perova
| 1 |
We used a fluorescently tagged Rab11 because the anti-Rab11 antibodies were not sufficient to label recycling endosomes. However, IFITM3 is detected at native levels and this compromise allowed us to evaluate its localization. Overexpressed IFITM3 was only used for the live cell microscopy shown in the movie, which has now been removed (see reply to reviewer 1).
| 2 | 1 |
Figure 7: I am not sure that this figure is a useful addition to the paper as it suffers from exactly the problems that the authors claim to overcome in their study, i.e., the use of overexpressed IFITM3 and an overexpressed Rab11 marker protein, both of which may show unnatural localization due to the overexpression.
| 1 | 2 |
v11060548_perova
| 1 |
Based on the reviewer´s suggestion, we have now included again the reference showing the infection increase of IAV in A549 IFITM3 knockdown cells and have changed the wording to make the statement clearer. “The mechanism by whichIFITM3 impedes IAV infection was divergent in previous studies and inhibition was mostly described upon IFITM3 over-expression [37, 38, 51]. It was demonstrated that IFITM3 knockdown in A549 cells increases infection rate [37].” Since this paper is primarily studying IFITM localization and clustering at endosomes/lysosomes it should reference studies in which endocytic localization motifs of the IFITMs were identified and characterized (Jia, Cell Microbiol, 2014; Chesarino, JBC, 2015; Li, JBC, 2015)
| 2 | 1 |
Lines 227/228: This sentence is misleading as the references cited are not contradictory, nor are they unclear as to IFITM3 possessing antiviral activity. Furthermore, the authors need to examine the literature more carefully, as previous studies have indeed shown that IFITM3 knockdown in A549 cells increases influenza virus infection, e.g., Lin, Cell Reports, 2013.
| 1 | 2 |
v11060548_perova
| 1 |
All the references have been added and we now include this aspect of the IFITM3 mechanism in the discussion of the paper. We thank the reviewer for the comment.
| 2 | 1 |
Since this paper is primarily studying IFITM localization and clustering at endosomes/lysosomes it should reference studies in which endocytic localization motifs of the IFITMs were identified and characterized (Jia, Cell Microbiol, 2014; Chesarino, JBC, 2014; Li, JBC, 2015) Line 65 states that subsequent studies have failed to confirm an association between SNP rs12252 and severe influenza.
| 1 | 2 |
v11060548_perova
| 1 |
Lines 61 ff. were changed pointing out the relevance of SNP rs12252-C and SNP rs34481144-A. References have been added.
| 2 | 1 |
Line 65 states that subsequent studies have failed to confirm an association between SNP rs12252 and severe influenza. This may be true, but far more studies have confirmed this link than have refuted it. Further, the studies which failed to find an association were generally performed in populations in which the SNP is almost non-existent. Additionally, a second SNP in the IFITM3 promoter has also been linked to severe flu. For a review of these studies, see Zani, Current Clin Microbiol Reports, 2018, though the primary articles should be cited.
| 1 | 2 |
v11060548_perova
| 1 |
We thank the reviewer for pointing out the inaccuracy in the references. The statement "...IFITM3 elevates the level of cholesterol on late endosomes and lysosomes thereby restricting early IAV infection" is based on the publication of Kühnl et al. 2018 (mBio). The reference was replaced. We have also complemented the introductory part with the aspect of fusion inhibition through the amphipathic helix of IFITM3.
| 2 | 1 |
Lines 67-73 The inhibition of virus membrane fusion by IFITM3 has been attributed to the presence of a palmitoylated amphipathic helix within IFITM3 (Chesarino, EMBO Reports, 2017). This helix and neighboring palmitoylation sites are among the most highly conserved residues among IFITMs from all species. Additionally, the role of cholesterol in IFITM3’s mechanism of action has been largely disproven by the field, and reference 52 seems to have been misused in line 73.
| 1 | 2 |
v11060548_perova
| 1 |
The innovation of this research has been clarified in the abstract, the introduction, and the conclusion.
| 2 | 1 |
The innovation of this research needs to be better explored.
| 1 | 2 |
w14030367_perova
| 1 |
Three references have been cited. Please see lines 40-41 in the revised manuscript.
| 2 | 1 |
Cite the following references: Panagopoulos, A. (2021). Energetic, economic and environmental assessment of zero liquid discharge (ZLD) brackish water and seawater desalination systems. Energy Conversion and Management, 235. Techno-economic assessment of Minimal Liquid Discharge (MLD) treatment systems for saline wastewater (brine) management and treatment. Process Safety and Environmental Protection, 146, pp. 656-669. Study and evaluation of the characteristics of saline wastewater (brine) produced by desalination and industrial plants. Environmental Science and Pollution Research, 1-14. Lines 33-38: You should mention that discharge of PPCPs degrades water quality and thus it cannot be directly used for potable water and industrial applications.
| 1 | 2 |
w14030367_perova
| 1 |
Thanks very much for the reviewer. The conclusion has been revised according to the reviewer’s suggestion. The added paragraph in the revised manuscript is as follow: To sum up, the results show that the proposed BAC-UF system can be effective in the treatment of river water polluted by PPCPs, conventional organic pollutants and ammonia nitrogen. Besides, the results of this analysis can have significant implications for the conventional UF operation procedure and the ozone-activated carbon process, providing a simple decentralized approach to drinking water treatment for the areas where source water is contaminated with PPCPs.
| 2 | 1 |
Conclusion: Discuss the applicability of your findings/results and future study in this field.
| 1 | 2 |
w14030367_perova
| 1 |
Thanks for the reviewer’s suggestion. The conclusion has been revised and integrated into two paragraphs. The first paragraph mainly includes important findings, and the second paragraph mainly includes the outlook for the future in this field.
| 2 | 1 |
Conclusion: Make it as one or two paragraphs.
| 1 | 2 |
w14030367_perova
| 1 |
The salinity concentrations in the samples have been added to Table 1 in the revised manuscript.
| 2 | 1 |
Table 1: What is the salinity (in mg/L) of the samples ?
| 1 | 2 |
w14030367_perova
| 1 |
We agree with the suggestion and comments of the reviewer. All comments have been revised one by one, and the modifications are highlighted in yellow in the marked revised manuscript.
| 2 | 1 |
However, there are some mistakes in the writing of the paper.
| 1 | 2 |
w14030367_perova
| 1 |
Thanks for the reviewer’s helpful suggestion. The objective statement has been added to the abstract to support the seriousness of the problem, and the revised sentences are as follows: Biological activated carbon (BAC) biofilter coupling ultrafiltration (UF) is a promising process for the treatment of river water contaminated by pharmaceutical and personal care products (PPCPs). However, the pilot-scale study should be conducted to reveal the long-term removal performance and the respective contributions of BAC and UF.
| 2 | 1 |
The title seems good, but the abstract seems to be fine. Please add one problem statement line in abstract to justify this sentence ``the long-term pilot-scale study is urged to be investigated.``.
| 1 | 2 |
w14030367_perova
| 1 |
We agree with the suggestion of this reviewer. The two-stage biofilms located in the activated carbon column and on the UF membrane synergistically, can be conducive to the removal performances. However, the mechanisms of the two-stage biofilm, such as bacterial and metazoan communities, membrane fouling and dissolved oxygen transfer, should be further investigated to enhance the removal efficiency and stability of this system. The research gap has been added in the Conclusion and is as follow: To sum up, the results show that the proposed BAC-UF system can be effective in the treatment of river water polluted by PPCPs, conventional organic pollutants and ammonia nitrogen. Besides, the results of this analysis can have significant implications for the conven-tional UF operation procedure and the ozone-activated carbon process, providing a simple decentralized approach to drinking water treatment for the areas where source water is contaminated with PPCPs.
| 2 | 1 |
Research gap should be delivered on more clear way with directed necessity for the future research work.
| 1 | 2 |
w14030367_perova
| 1 |
Thanks for the reviewer’s comment. The introduction has been revised carefully, including using up-to-date references. For example, the outdated references have been removed. Meanwhile, the articles published within the last two years were added, such as (the Reference2 Yu et al., 2020) and (the Reference21 Tang et al., 2018).
| 2 | 1 |
Introduction section must be written on more quality way, i.e., more up-to-date references addressed. Please target the specific gap such as 2015-2021 etc.
| 1 | 2 |
w14030367_perova
| 1 |
The reference has been cited. Please see line 38 in the revised manuscript.
| 2 | 1 |
Page 1 Line 38. Please cite this reference with existing reference 3….Role of nanotechnology for design and development of cosmeceutical: application in makeup and skin care.
| 1 | 2 |
w14030367_perova
| 1 |
The BAC biofilter can remove the PPCPs, and then the following UF can reject micro-organisms and particles flowing out from the biofilter to ensure the quality of drinking water. The above coupling process makes up for the defects concerning respective operations of the BAC biofilter and UF. Although many pilot-scale setups were used to treat secondary wastewater effluent for water reclamation, this type of raw water quality was different from the river water, causing the different potential of biofilm growing. As far as we investigated, the lack of enough attention to long-term pilot study exists. Therefore, in this study, a BAC-UF system was carried out for several months with pilot scale to access the long-term removal performances and the respective contributions of BAC and UF. The relevant content has been supplemented and revised in the abstract to highlight the innovative points.
| 2 | 1 |
The novelty of the work must be clearly addressed and discussed, compare previous research with existing research findings and highlight novelty.
| 1 | 2 |
w14030367_perova
| 1 |
Biological activated carbon (BAC) combined the adsorption and biologic degradation consuming low power energy and chemicals without concern of DBPs production as well as no frequent updates for activated carbon media. The BAC biofilter can remove the PPCPs pollutants, and then the followed UF can reject microorganisms and particles flowing out from the biofilter to ensure the quality of drinking water. Thus, the BAC-UF process makes up for the defects of the respective operations of BAC biofilter and UF. Although many pilot-scale setups were used to treat secondary wastewater effluent for water reclamation, this type of raw water quality was different from the river water, causing the potential of biofilm growing differently. As far as we investigated, the lack of enough attention to long-term pilot study is present. Therefore, in this study, a BAC-UF system was carried out for several months with a pilot scale.
| 2 | 1 |
What is the main challenge? Why author choose this material? Please highlight in the introduction part.
| 1 | 2 |
w14030367_perova
| 1 |
The two references have been cited. Please see the lines 58.
| 2 | 1 |
Page 2 Line 55 need a reference. Please consider these at end of this sentence……The oxidation method exhibited a fast reaction speed and high removal efficiency…(i) Role of nanomaterials in the treatment of wastewater: A review (ii) Advances and challenges in developing efficient graphene oxide-based ZnO photocatalysts for dye photo-oxidation.
| 1 | 2 |
w14030367_perova
| 1 |
The introduction has been improved for better reading.
| 2 | 1 |
The main objective of the work must be written on the more clear and more concise way at the end of introduction section.
| 1 | 2 |
w14030367_perova
| 1 |
Two PPCP mistakes have been revised. The abbreviation of chemical oxygen demand in previous version has been corrected to CODMn. The unnecessary abbreviations have been also corrected in the abstract.
| 2 | 1 |
Please check the abbreviations of words throughout the article. All should be consistent.
| 1 | 2 |
w14030367_perova
| 1 |
The added paragraph is as follows: Potassium permanganate, H2SO4, potassium sodium tartrate tetrahydrate, Nessler’s reagent and NaOH were purchased from a commercial company and certified as AR purity (Guangzhou Chemical Reagent Factory, Guangzhou, China), while PPCP standards were provided by three companies. Sulfamethoxazole (SMX), Sulfadoxine (SD), Sulfachloropyridazine (SCP), Sulfamethoxypyridazine (SMP), Sulfaquinoxaline (SQX), Sulfathiazole (STZ), Doxycycline (DOX), Erythromycin (EM), Anhydroerythromycin (EA), Roxithromycin (ROX), Penicillin-G (PEN G), Clarithromycin (CAM), Norfloxacin (NOR), Oxociprofloxacin (OFL), Enrofloxacin (EFL), Flumequine (FQ), Acetaminophen (APAP), Diclofenac sodium (DCF), Naproxen (NAP), Indomethacine (IND), Metoprolol (METO), Propranolol (), Atenolol (ATL), Primidone (PRM), Carbamazepine (CMZ), and Sulpiride (SP) were obtained from the Dr. Ehrenstorfer Company in Germany. Furthermore, Sulfadimidine (SM2), Sulfadiazine (SDZ), Sulfapyridine (SP), Sulfamonomethoxine (SMM), Tetracycline (TC), Ofloxacin (OFL), Amoxicillin (AM), Dimetridazole (DMZ), Trimethoprim (TMP) were bought from the National Institute of Metrology in China, whereas Oxytetracycline (OTC), Caffeine (CF), and Diethyltoluamide (DEET) were acquired from the Toronto Research Chemicals Company in Canada.
| 2 | 1 |
Please add chemical reagents section and stated all chemical with brand specifications.
| 1 | 2 |
w14030367_perova
| 1 |
These three GAC-UF systems were operated in parallel. Samples of the feedwater and effluent from three systems were taken simultaneously and measured once. The standard deviation was obtained by the detection over the full period of the experiment.
| 2 | 1 |
Regarding the replications, authors confirmed that replications of experiment were carried out. However, these results are not shown in the manuscript, how many replicated were carried out by experiment? Results seem to be related to a unique experiment. Please, clarify whether the results of this document are from a single experiment or from an average resulting from replications. If replicated were carried out, the use of average data is required as well as the standard deviation in the results and figures shown throughout the manuscript. In case of showing only one replicate explain why only one is shown and include the standard deviations.
| 1 | 2 |
w14030367_perova
| 1 |
The figure is used as the only pattern in the revised manuscript.
| 2 | 1 |
Please revise your paper accordingly since some issue occurs on several spots in the paper. Please use Fig. or figure? It very confusing. Article should be in one pattern.
| 1 | 2 |
w14030367_perova
| 1 |
The front sizes in the figure 1 have been increased. The quality of the figure 1 has been also improved.
| 2 | 1 |
Please provide high quality image of figure
| 1 | 2 |
w14030367_perova
| 1 |
The style for units has been revised and unified.
| 2 | 1 |
Please use one style for units such as m3/h or m3h-1 Please revise your paper accordingly since some issue occurs on several spots in the paper.
| 1 | 2 |
w14030367_perova
| 1 |
The comparison of previous research with existing research findings was added in the final part of the results and discussion. The added paragraph is as follows: PPCPs risks have been posing severe challenges to the safety of drinking water supply in rural areas due to the absence of the process with simple operation and maintenance as well as reliable performance. In this study, BAC coupling gravi-ty-driven UF was performed continuously, and the rejection performance of mem-brane filtration and BAC filtration both showed barriers for the conventional pollu-tants and PPCPs. Furthermore, this study indicated the respective contributions of BAC and UF, showing the role of the two-stage biofilm. Previous works involving BAC generally combined the ozonation with the BAC filter for treating the contaminants of emerging concern, eliminating a majority of PPCPs by more than 90% [34]. However, the regulation and maintenance of machines for ozone products are complicated, and the disinfection by-products will be newly generated in the effluent, which is incon-venient to use in rural areas [35]. In general, coagulation, filtration and single BAC units worked inefficiently and removed the detected PPCPs by less than 50%, as they were not hydrophobic [34, 36]. Hybrid membrane processes such as inline dosing of powdered activated carbon (PAC) prior to UF have already shown promising potential for the abatement of PPCPs; however, the inline dosing PAC is infeasible in rural areas [37]. In this study, the BAC prior to UF enhanced the biological activity by forming a two-stage biofilm system. Therefore, the integrated BAC-UF process can be considered as an economically and technically feasible approach to the decentralized and emer-gency drinking water treatment.
| 2 | 1 |
Please add a comparative profile section to compare your results and prove how it better than previous.
| 1 | 2 |
w14030367_perova
| 1 |
The conclusion has been modified as suggested by reviewers. Please see lines 325-350 in the revised manuscript.
| 2 | 1 |
Section 4 should be renamed by Conclusion and Future perspectives. Conclusion section is missing some perspective related to the future research work, quantify main research findings, highlight relevance of the work with respect to the field aspect. In the present form conclusion is very weird.
| 1 | 2 |
w14030367_perova
| 1 |
Two native English-speaking colleagues help us verify the manuscript. Hope the revised manuscript would be more satisfactory. We are so sorry to make reviewer’s reading uncomfortable. We have used an English Language Editing service to correct the grammatical and spelling errors and to make the expressions conform to correct scientific English (the Language Editing Certification is attached below).
| 2 | 1 |
To avoid grammar and linguistic mistakes, Major level English language should be thoroughly checked. Please revise your paper accordingly since several language issue occurs on several spots in the paper.
| 1 | 2 |
w14030367_perova
| 1 |
We agree with the reviewer. The reference formatting has been corrected. Thanks very much for the patient review again.
| 2 | 1 |
Please follow the journal guidelines. Reference formatting need carefully revision. All must be consistent in one formate.
| 1 | 2 |
w14030367_perova
| 1 |
Full names of BAC, UF, and PPCPs are used in the title and the abstract to show the results briefly. As for NH4+-N, NO2−-N, and NO3—N, the full names have been used instead of abbreviations in the abstract.
| 2 | 1 |
Authors should avoid abbreviations in the title and the abstract.
| 1 | 2 |
w14030367_perova
| 1 |
The title has been revised to river water.
| 2 | 1 |
Authors may revise the title to include river water instead of surface water.
| 1 | 2 |
w14030367_perova
| 1 |
The references format has been corrected.
| 2 | 1 |
References should be according to the journal format.
| 1 | 2 |
w14030367_perova
| 1 |
We are very sorry for the confusion caused to the reviewers, the sentences have been revised from the words and Grammarly. The revised sentences are shown below: Ultrafiltration (UF) as an emerging alternative technology to conventional water treatment processes, has been widely used to remove pollutants such as particles, col-loids, bacteria, and viruses, thus reducing the risk of water-borne diseases [10]. However, in the case of the PPCPs with a small molecular weight (typically < 600 Da), UF membranes also cannot effectively reject these PPCPs, but nanofiltration and reverse osmosis are able to remove these PPCPs based on the thin-film composite [11, 12].
| 2 | 1 |
Line 44-45: Ultrafiltration (UF) as emerging technology, has been widely used to remove pollutants such as particles, colloids, bacteria and viruses, reducing the risk of water-borne diseases and …UF membranes cannot effectively rejected these soluble substances”. Please clarify, why ultrafiltration cannot remove the PPCPs since it can remove bacteria and viruses.
| 1 | 2 |
w14030367_perova
| 1 |
The t-test results have been added in the revised Table1.
| 2 | 1 |
Please add t-test results in Table 1 for each parameter to understand the significant differences. Authors may provide data in the supporting information file.
| 1 | 2 |
w14030367_perova
| 1 |
The statistical t-test was evaluated for Figure 4, which was added in the text of "3.2. Removal of nitrogen" Section.
| 2 | 1 |
The reviewer suggests evaluating the statistical t-test for Figures 4.
| 1 | 2 |
w14030367_perova
| 1 |
The t-test results have been added to the revised Table2. The concentration of Erythromycin was generally varied between 1047.14 ng L−1 and 2037.72 ng L−1.
| 2 | 1 |
t-test results should be included in Table Why is the standard deviation of Erythromycin showing a high value?
| 1 | 2 |
w14030367_perova
| 1 |
Some symbols were overlapped in Figure 5h. The symbols near 85 days have been entirely shown by correcting Y-axis settings, such as Figures 5a and 5f.
| 2 | 1 |
Please look at the curve of BAC/UF; there was a symbol missing in near 85 days. Please show each symbol.
| 1 | 2 |
w14030367_perova
| 1 |
The conclusion has been revised, including the style and sentences.
| 2 | 1 |
The conclusion may be revised.
| 1 | 2 |
w14030367_perova
| 1 |
We agree with this suggestion. The abbreviations in the title have been deleted. Besides, a list of abbreviations has been added before the references.
| 4 | 1 |
Authors should avoid abbreviations in the title. Authors should add a list of abbreviations before the references.
| 3 | 2 |
w14030367_perova
| 1 |
Thanks to the reviewer for the patient comment. The above mistakes have been revised.
| 4 | 1 |
Please avoid repeating the full name and abbreviation throughout the manuscript if you used the first-time abbreviation.
| 3 | 2 |
w14030367_perova
| 1 |
1. 2. Thanks to this reviewer for the comment. Common waterborne viruses include Shigella, Vibrio cholerae, Poliovirus, adenovirus, and coxsackie virus, with sizes of 26 nm, 27 nm, 30 nm, 90 nm, and 30 nm, respectively [1-5]. The molecular weights of PPCPs are generally less than 1000 Da (about 1-2 nm), obviously smaller than the virus. The molecular weight cut off of UF membranes was an average 100 000 Da in this work, which was similar to the virus but larger than the PPCPs molecules. Karthik K, Dhanuskodi S, Gobinath C, et al. Multifunctional properties of microwave assisted CdO–NiO–ZnO mixed metal oxide nanocomposite: enhanced photocatalytic and antibacterial activities [J]. Journal of Materials Science: Materials in Electronics, 2018, 29(7): 5459-71. Song S, Liu Z, Zhou J, et al. An adjuvant compound that enhances immunogenicity at fractional doses of the Sabin-inactivated poliovirus vaccine (sIPV) with a long duration of protection in a rat model [J]. Journal of Medical Virology, 2019, 91(1): 14-21. Kim K, Choi J-W, Ma K, et al. Nanoisland-Based Random Activation of Fluorescence for Visualizing Endocytotic Internalization of Adenovirus [J]. Small, 2010, 6(12): 1293-9. Dourmashkin R R, Mccall S A, Dourmashkin N, et al. Virus-like particles and enterovirus antigen found in the brainstem neurons of Parkinson's disease [J]. F1000Res, 2018, 7: 302-. Tamano K, Aizawa S-I, Katayama E, et al. Supramolecular structure of the Shigella type III secretion machinery: the needle part is changeable in length and essential for delivery of effectors [J]. The EMBO Journal, 2000, 19(15): 3876-87.
| 4 | 1 |
Line 46-52: & line 84-85 “Ultrafiltration (UF) as an emerging alternative technology to conventional water treatment processes, has been widely used to remove pollutants such as particles, colloids, bacteria, and viruses, thus reducing the risk of water-borne diseases [10]. Size exclusion is considered the primary removal mechanism for the UF. However, in the case of the PPCPs with a small molecular weight (typically < 600 Da), UF membranes also cannot effectively reject these PPCPs, but nanofiltration and reverse osmosis are able to remove these PPCPs based on the thin-film composite”. Sorry, but I don't see why UF can reject viruses and bacteria but not PPCPs. Is it true that the molecular weight of viruses and bacteria is more than that of PPCPs? Please elaborate.
| 3 | 2 |
w14030367_perova
| 1 |
The information regarding the biological degradation of PPCPs has been added in the introduction. The revised sentences are shown below: In the BAC biofilters, the biotransformation and adsorption both contributed to the PPCPs removal. The activated carbon adsorbed PPCPs to the surface and interior, where microorganisms were suitable for growth. Under the long-term effect of high-concentration PPCPs, the dominant microorganisms in the biofilter were selected to survive. These microorganisms mostly transformed PPCPs into many segments and even directly mineralized them to CO2 [3, 14].
| 4 | 1 |
The authors should include some information regarding the biological degradation of PPCPs by microorganisms in the introduction section.
| 3 | 2 |
w14030367_perova
| 1 |
“The combined process with ultrafiltration may be another promising choice, featuring a comparable removing performance as the nanofiltration and low operational cost”. This claim is not supported by our research. To avoid misunderstandings for readers, we changed the claim and revised the sentence. The revised version is as follows: The combined process with UF may be another promising choice as an alternative to nanofiltration for removing PPCPs in rural areas. It can be seen in lines 54-55.
| 4 | 1 |
Line 53-55: Have the authors compared the BAC-UF performance to that of nanofiltration?
| 3 | 2 |
w14030367_perova
| 1 |
The method of the t-tests has been explained in section 2.4 (lines 157-158) in the revised manuscript. The sample size (n) has also been added in Table 1 and Table 2.
| 4 | 1 |
I strongly suggest that the authors explain how they performed the t-tests. Please include the sample size (n) or degree of freedom (df).
| 3 | 2 |
w14030367_perova
| 1 |
Thank you very much for your kind reminder. The values have been modified in the revised manuscript in yellow highlight.
| 4 | 1 |
Please show the t-value and p-value in a scientific view (e.g., 2.79E-4 would be P <.001).
| 3 | 2 |
w14030367_perova
| 1 |
Thanks for the kind reminder. All the names of medicinal compounds not used have been deleted, including Sulfaquinoxaline (SQX), Sulfathiazole (STZ), Doxycycline (DOX), Roxithromycin (ROX), Penicillin-G (PEN-G), Clarithromycin (CAM), Norfloxacin (NOR), Oxociprofloxacin (OFL), Enrofloxacin (EFL), Flumequine (FQ), Acetaminophen (APAP), Diclofenac sodium (DCF), Naproxen (NAP), Indomethacin (IND), Metoprolol (METO), Propranolol (PRO), Atenolol (ATL), Primidone (PRM), Carbamazepine (CMZ), Sulpiride (SP), Sulfapyridine (SPN), Sulfamonomethoxine (SMM), Tetracycline (TC), Amoxicillin (AM), Dimetridazole (DMZ), Oxytetracycline (OTC) and DEET.
| 4 | 1 |
Line 161-176: Please delete the name of medicinal compounds not used for this study.
| 3 | 2 |
w14030367_perova
| 1 |
The results and discussion has been supplemented in section 3.1. Please see lines 180-185 in the revised manuscript.
| 4 | 1 |
1) was observed in Fig. 1(c). Why is the DO concentration of BAC-effluent sometimes higher than the Raw water. Please explain it in the manuscript. The reviewer suggests statistical analysis using a t-test (Raw water- BAC-Effluent and Raw water – BAC/UF-effluent). Point 6: Why the same trend of increase or decrease in graphs (Fig. 1) was observed in Fig. 1(c).
| 3 | 2 |
w14030367_perova
| 1 |
Thanks very much for this comment. With a turbulent current and dozens of kilometers in length, this river is located in the mountains of Foshan city. The PPCPs sludge at the bottom of the river never executes removal. Thanks for the suggestion from the reviewer. We will further precipitate the PPCPs sludge before the water treatment plant or excavate the PPCPs sludge from the river and study the effects on drinking water quality. The relevant results will be published in another paper in the future.
| 4 | 1 |
The same trend (Figure 2) was mainly due to the stable removal ability of BAC and UF for organics, causing the removal restriction. The periodic backwash (7days) of BAC caused sometimes the dissolved oxygen concentration of BAC-effluent higher than that of raw water. After the gas scrubbing and the hydraulic backwashing, the dissolved oxygen detection of the effluent was carried out, resulting in the above results for dissolved oxygen. Besides, the t-test was used and proved the significant difference between BAC-Effluent and BAC/UF-effluent. Response 6: Thanks very much for this comment.
| 3 | 2 |
w14030367_perova
| 1 |
Accepted and revised in L95-105. Five-point sampling method was used in both fields (0-20cm). A total of 5 topsoil samples in each field were taken using a stainless steel spade and mixed thoroughly. Afterwards the soil samples were chosen by quartation; Then plants’ debris and residues were removed. Finally soil samples were 0.3-mm sieved and stored in brown glass bottles at 4 °C. The following was the area that the Florida soil was collected.
| 2 | 1 |
There is no sampling procedure, depth, or random selection of the soils evaluated.
| 1 | 2 |
w14081258_makarova
| 1 |
It has been mentioned in L163-164 in the manuscript. Randic (1975) proposed molecular branching index χ, hereinafter referred to as simple molecular connectivity index (1995). Therefore, they are the same concept. [38] Randic M., Razinger M. (1995). Molecular Topograhic Indices. Journal of Chemical Information and computer Sciences. 35: 140-147. [40] Randic M. (1975). On Characterization of molecular branching. Am. Chem. Soc. 97: 6609-6615. The values obtained of I DW and steric hindrance are not explained.
| 2 | 1 |
Is the explanation of χ is for molecular connectivity indices or branching index?
| 1 | 2 |
w14081258_makarova
| 1 |
The analytical methodology of isomers quantification has been studied and published in former study (Wang et al., 2013). From Figure1 in Supporting Information, it seems that the isomers were not separated, but actually it does not affect the quantification of the isomers. Because the main quantitative ions were used for quantification and the main quantitative ion of each isomer were separated absolutely (Figure 1). Table 1 and Figure 1 were cited from Wang et al (2013). Moreover, series of research about the NP isomers were conducted and published afterwards. The following are the publications. Shiyu Wang, Fei Liu*, Wenyong Wu*, Yaqi Hu, Renkuan Liao, Gaoting Chen, Jiulong Wang, Jialin Li. Migration and health risks of nonylphenol and bisphenol A in soil-winter wheat systems with long-term reclaimed water irrigation. Ecotoxicology and Environmental Safety 158 (2018) 28–36. Shiyu Wang，Wenyong Wu*，Fei Liu, Xiaoou Li. Sorption and desorption behaviours of 4-nonylphenol on reclaimed water-irrigated soils. Environmental Engineering Science. (2019) 36 (9) : 1100-1111。 王世玉，刘菲*，刘玉龙，陈亮. 气相色谱-质谱法检测地下水中12种对壬基酚同分异构体.分析化学，2013，41（11）：1699-1703. 王世玉，刘菲*，吴文勇，尹世洋，刘玉龙，陈亮，张伟，陈会会.影响12种壬基酚同分异构体液液萃取效率的因素研究. 岩矿测试，2014，33（4）：570-577.
| 2 | 1 |
If correct quantification is not assessed, then figure 1 and table 1 and 2 are not valid.
| 1 | 2 |
w14081258_makarova
| 1 |
It has been given in Supporting Information 2. Indeed, the characteristics of the soil are too limited. My visiting time was only one year. At the end of the visiting time, I had no time to complete the microbiological test and the soil properties detection and was going to leave the United States. So I had to acquire the Florida soil properties information by website. However, the information I can acquire from the website was limited, which were shown in Supporting Information 2.
| 2 | 1 |
The basic physi-chemical parameters of soils should be given.
| 1 | 2 |
w14081258_makarova
| 1 |
Shiyu Wang, Wenyong Wu*, Fei Liu, Shiyang Yin, Zhe Bao, Honglu Liu. Spatial distribution and migration of nonylphenol in groundwater following long-term wastewater irrigation. Journal of Contaminant Hydrology,2015,177-178（June–July）:85-92. I think it is a little difficult for me to acquire the irrigation water physi-chemical parameters of Florida, but I can acquire the NP load in China irrigation water, which were cited from my former study of Wang et al., (2015). Bu in this study, the irrigation water was prepared in Lab according to the max NP concentration in reclaimed water in the study area to simulate NP in reclaimed water in the actual environment.
| 2 | 1 |
The irrigation water physi-chemical parameters should be given.
| 1 | 2 |
w14081258_makarova
| 1 |
It has been given in Table1.
| 2 | 1 |
The kinect models should be given.
| 1 | 2 |
w14081258_makarova
| 1 |
Accepted and revised in Figure1. All the isomers were revised according to kinect model except some isomers, such as NP2 NP5 and NP11. The degradation of these isomers were stable within the former several days, which mentioned in the manuscript. So these former points conformed to first-order kinetic formula.
| 2 | 1 |
Figure 1 should be revised according to kinect model.
| 1 | 2 |
w14081258_makarova
| 1 |
The main reason is that this experiment was conducted during my being in University of Florida, USA as a visiting scholar. Worse more, the soil can not be brought to China. Indeed, in this study, the analysis of biomass and microorganism of the soil samples is essential. But I did not do that. My visiting time is only one year. At the end, I had no time to complete and analysis the microbial community and biomass of the soil samples. But in the further study of the NP isomers in reclaimed water soil, this should be taken into account.
| 2 | 1 |
I strongly recommend the authors to analysis the biomass of the soil samples.
| 1 | 2 |
w14081258_makarova
| 1 |
Actually, the soil samples were 0.25 mm sieved. At first, the soil was 0.3 mm sieved, but the big particle size was not appropriate for the soil ultrasonic treatment of NP extraction. Therefore, 0.3mm-sieved soil was not used for the experiment(which has been deleted in the manuscript in L104). Then the soil was 0.25mm-sieved, which was suitable for the NP extraction. If the soil was 2 mm-sieved, the extraction of the NP was more inappropriate. So 0.25mm sieve was used in this study. Indeed, this size particle excluded an important part of the active soil where microorganisms live and organic carbon is accumulated. But for this study, the degradation was completed within 30 days in both of the two soils. Therefore, though some of the organic carbon was adsorbed on the surface of the particle, the amounts of the microorganisms are enough for the degradation of NP. However, the degradation rate could be affected by this. In the future study, this would be taken into accounted. What’s more, the microbial community could be taken into accounted as well. Accepted.
| 4 | 1 |
I do not understand why soil samples were 0.3 mm sieved. Why? The active soil components (lime, sand, clay) where microorganisms live and organic carbon is accumulated is just less than 2mm. Consequently you excluded an important part of the active soil from your study, therefore your resuts are very limited.R： Moreover, in the subsequent sentence is reported: "The soil was 0.25 mm-sieved to remove large particles; and then weighed series of 10 g aliquots into 250 mL brown jars" Did you sieve the soil twice at 0.3 mm and then 0.25 mm?
| 3 | 2 |
w14081258_makarova
| 1 |
Accepted and revised in L204. Figure 1 has been changed in the manuscript.
| 4 | 1 |
Figure 1 legends of axis need font size larger, please make readable the figuresR：
| 3 | 2 |
w14081258_makarova
| 1 |
Accepted and revised in L95-105. Five-point sampling method was used in both fields (0-20cm). A total of 5 topsoil samples in each field were taken using a stainless steel spade and mixed thoroughly. Afterwards the soil samples were chosen by quartation; Then plants’ debris and residues were removed. Finally soil samples were 0.3-mm sieved and stored in brown glass bottles at 4 °C. The following was the area that the Florida soil was collected.
| 2 | 1 |
There is no sampling procedure, depth, or random selection of the soils evaluated.
| 1 | 2 |
w14081258_perova
| 1 |
It has been mentioned in L163-164 in the manuscript. Randic (1975) proposed molecular branching index χ, hereinafter referred to as simple molecular connectivity index (1995). Therefore, they are the same concept. [38] Randic M., Razinger M. (1995). Molecular Topograhic Indices. Journal of Chemical Information and computer Sciences. 35: 140-147. [40] Randic M. (1975). On Characterization of molecular branching. Am. Chem. Soc. 97: 6609-6615.
| 2 | 1 |
Is the explanation of χ is for molecular connectivity indices or branching index?
| 1 | 2 |
w14081258_perova
| 1 |
Accepted and revised in L157-160. Common topological indices, such as κ shape indices and information indices were also calculated by Molconn-Z (version 4.12S, eduSoft, La Jolla, CA). The detailed list and definitions can be found in the software user’s guide [35], and the definitions can also be found in Todeschini et al. [36]. The results of other common topological indices calculated by Molconn-Z were shown below as well. But in this study, only the steric index (R2 = 0.82 for CN soil; R2 = 0.86 for FN soil) and IDWbar (R2 = 0.83 for CN soil; R2 = 0.94 for FN soil) have a better relationship with the half lives of the isomers, which were mentioned in the Results and Discussion part. The results of the topological indices of the isomers [35] Hall, L. H.; Kellogg, G. E.; Haney, D. N. (2008). Software Package for Molecular Topology Analysis User’S Guide. [36] Todeschini, R.; Consonni, V.; Mannhold, R.; Kubinyi, H.; Folkers, G. (2009). Molecular Descriptors for Chemoinformatics; Wiley: New York.
| 2 | 1 |
The values obtained of I DW and steric hindrance are not explained.
| 1 | 2 |
w14081258_perova
| 1 |
The analytical methodology of isomers quantification has been studied and published in former study (Wang et al., 2013). From Figure1 in Supporting Information, it seems that the isomers were not separated, but actually it does not affect the quantification of the isomers. Because the main quantitative ions were used for quantification and the main quantitative ion of each isomer were separated absolutely (Figure 1). Table 1 and Figure 1 were cited from Wang et al (2013). Moreover, series of research about the NP isomers were conducted and published afterwards. The following are the publications.
| 2 | 1 |
The more critical issue lies in the analytical methodology used for isomers quantification. First of all, figure one is a chromatogram, not a spectrum. The chromatogram is not well resolved, i.e., there is no separation among the different analytes determined. For example, look at peaks 4, 5, and 6. There is no resolution among the isomers, the same for NP 7 and 8 and NP 8 and 10; thus, it is impossible to quantify them. If correct quantification is not assessed, then figure 1 and table 1 and 2 are not valid.
| 1 | 2 |
w14081258_perova
| 1 |
It has been given in Supporting Information 2. Indeed, the characteristics of the soil are too limited. My visiting time was only one year. At the end of the visiting time, I had no time to complete the microbiological test and the soil properties detection and was going to leave the United States. So I had to acquire the Florida soil properties information by website. However, the information I can acquire from the website was limited, which were shown in Supporting Information 2.
| 2 | 1 |
The basic physi-chemical parameters of soils should be given.
| 1 | 2 |
w14081258_perova
| 1 |
I think it is a little difficult for me to acquire the irrigation water physi-chemical parameters of Florida, but I can acquire the NP load in China irrigation water, which were cited from my former study of Wang et al., (2015). Bu in this study, the irrigation water was prepared in Lab according to the max NP concentration in reclaimed water in the study area to simulate NP in reclaimed water in the actual environment.
| 2 | 1 |
The irrigation water physi-chemical parameters should be given.
| 1 | 2 |
w14081258_perova
| 1 |
It has been given in Table1.
| 2 | 1 |
The kinect models should be given.
| 1 | 2 |
w14081258_perova
| 1 |
Accepted and revised in Figure1. All the isomers were revised according to kinect model except some isomers, such as NP2 NP5 and NP11. The degradation of these isomers were stable within the former several days, which mentioned in the manuscript. So these former points conformed to first-order kinetic formula.
| 2 | 1 |
Figure 1 should be revised according to kinect model.
| 1 | 2 |
w14081258_perova
| 1 |
The main reason is that this experiment was conducted during my being in University of Florida, USA as a visiting scholar. Worse more, the soil can not be brought to China. Indeed, in this study, the analysis of biomass and microorganism of the soil samples is essential. But I did not do that. My visiting time is only one year. At the end, I had no time to complete and analysis the microbial community and biomass of the soil samples. But in the further study of the NP isomers in reclaimed water soil, this should be taken into account.
| 2 | 1 |
I strongly recommend the authors to analysis the biomass of the soil samples.
| 1 | 2 |
w14081258_perova
| 1 |
Actually, the soil samples were 0.25 mm sieved. At first, the soil was 0.3 mm sieved, but the big particle size was not appropriate for the soil ultrasonic treatment of NP extraction. Therefore, 0.3mm-sieved soil was not used for the experiment(which has been deleted in the manuscript in L104). Then the soil was 0.25mm-sieved, which was suitable for the NP extraction. If the soil was 2 mm-sieved, the extraction of the NP was more inappropriate. So 0.25mm sieve was used in this study. Indeed, this size particle excluded an important part of the active soil where microorganisms live and organic carbon is accumulated. But for this study, the degradation was completed within 30 days in both of the two soils. Therefore, though some of the organic carbon was adsorbed on the surface of the particle, the amounts of the microorganisms are enough for the degradation of NP. However, the degradation rate could be affected by this. In the future study, this would be taken into accounted. What’s more, the microbial community could be taken into accounted as well. Accepted.
| 2 | 1 |
I do not understand why soil samples were 0.3 mm sieved. Why? The active soil components (lime, sand, clay) where microorganisms live and organic carbon is accumulated is just less than 2mm. Consequently you excluded an important part of the active soil from your study, therefore your resuts are very limited.R： Moreover, in the subsequent sentence is reported: "The soil was 0.25 mm-sieved to remove large particles; and then weighed series of 10 g aliquots into 250 mL brown jars" Did you sieve the soil twice at 0.3 mm and then 0.25 mm?
| 1 | 2 |
w14081258_perova
| 1 |
Accepted and revised in L204. Figure 1 has been changed in the manuscript.
| 2 | 1 |
Figure 1 legends of axis need font size larger, please make readable the figuresR：
| 1 | 2 |
w14081258_perova
| 1 |
However, on pages 7 - 9 the authors admit that they divert from the CIRA procedure by identifying the outcome in advance, assigning qualitative values to preferences, assuming weights, utility factors, and initial values, and considering several stakeholders as risk owners.
| null | null |
As a separate recommendation, the paper needs smoother transitions between topics on pages 2-7. The authors should add one or two lines smoothing the flow for the reader, rather than relying on subject headings.
| 1 | 2 |
admsci5030125_boyarkin
| 0 |
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