Use of Biostimulants Obtained from Sewage Sludge for the Restoration of Soils Polluted by Diuron: Effect on Soil Biochemical Properties

Round 1

Reviewer 1 Report

The ms xx with the title of Use of biostimulants obtained from sewage sludge in the restoration of polluted soils by diuron. Effect on soil biochemical properties investigate an interesting topic. The authors investigated  the biorremediation effect of three biostimulants obtained from sewage slude through an enzymatic hydrolysis process and fermentation process in a diuron-contaminated soil for 55 days under laboratory conditions. However, the authors have to improve the ms before it can be accepted in such a high-quality journal. Below are my comments:

Affiliations: the authors should write their affiliations in English instead of Spanish.

The ms need significant improvements in terms of English

The authors have to reduce the self-citations, I noted that self-citation is about 50% of all cited references. I do not know how many authors can make self-citations per their work? Five? But not over than ten at least.

L14 manuscrpito? Is this a Spanish word? I think you mean manuscript! But, the word manuscript is not suitable to be used here, instead you can write (In this study, we …)

L14 add , after manuscrpito

L19 bacGram? What is this? I know it, but authors should define any abbreviations in the first mention to make the text clear and understandable for readers.

L20 what is Gram? Please define

L19-21 please add was before the word of decreased in this sentence. So, add it three times.

L22-23 greater than what? Please add full meaning for text.

L26 add sewage sludge as keyword

L33 remove these

L33-35 please cite these relevant ref: https://doi.org/10.1016/j.chemosphere.2012.07.031  

L35 or in other places: please use sludge as a singular instead of using this as plural

L37-42 Please cite the relevant references for this text.

L43-46 Please cite this very relevant ref: https://doi.org/10.1016/j.resconrec.2019.104647

https://doi.org/10.1016/j.chemosphere.2006.12.019

L47-49 please add this relevant citation: https://doi.org/10.1016/j.biombioe.2013.07.021

L55 what are these studies? Please cite two of the relevant or some of the above-cited references.

L91 was NOT is

L27 ha-1 make -1 as superscript

L130-131 based on what you added 9.23 mg for each 300 g soil to get 2 kg per ha? Then, how many kg soils per ha do you think?

L133 BS1, BS2 and BS3??? What are these? Suddenly appeared, please define

L157 why authors decided to analyze the diuron content in the soil at 2, 6, 10, 20, 35 and 55 days? Why was not done at 5,10,20,40,55 or …?

L163-170 revise the Statistical analysis, it was not well written.

Table 1: Why S, Ca and Pb are very high concentrations??

L201 please use p ≤0.05 NOT p <0.05 , correct this in other 9 places within your ms.

Table 2, can be converted to Figure?

Table 2: Why β-glucosidase activity was not reduced significantly as Dehydrogenase activity was reduced by increasing the period on incubation? The reason? This should be deeply explained in the discussion section.

Discussion section has to be significantly improved, the authors have to make it deeper and explain the different mechanisms of their treatments in reducing or increasing their parameters instead of saying that our results were in line with this and this. If you say this, what is the new of your investigation?

L343-344 common text, remove it. In conclusion, authors should focus on the most important finding at their work.

 

I am sure that authors will be able to improve their ms.

Regards,

Reviewer

 

 

 

 

 

 

 

 

 

Author Response

The ms xx with the title of Use of biostimulants obtained from sewage sludge in the restoration of polluted soils by diuron. Effect on soil biochemical properties investigate an interesting topic. The authors investigated  the biorremediation effect of three biostimulants obtained from sewage slude through an enzymatic hydrolysis process and fermentation process in a diuron-contaminated soil for 55 days under laboratory conditions. However, the authors have to improve the ms before it can be accepted in such a high-quality journal. Below are my comments:

 

Before beginning to answer the referees' questions, the authors want to comment on one matter.

Fortunately reviewer 1 made us a comment about the amount of soil used in the experiment and after reviewing the data we observed that we were confused with data from another experiment that we are carrying out with diuron.

In this experiment developed in this manuscript, the authors used 9.3 kg of soil for a dose of 2 kg/ha of diuron, with which 9.23 mg of the herbicide was added to this amount of soil.

We hope that this error of ours will not negatively influence the decision on the manuscript.

The authors want to make it clear that the data in the tables of the biological properties of the soil as well as those of the concentration of diuron in the soil are correct.

therefore we apologize for this great mistake that we have committed and that fortunately we have corrected

 

Affiliations: the authors should write their affiliations in English instead of Spanish.

Revised

 

The ms need significant improvements in terms of English

The manuscript has now been reviewed by a native English speaker

 

The authors have to reduce the self-citations, I noted that self-citation is about 50% of all cited references. I do not know how many authors can make self-citations per their work? Five? But not over than ten at least.

The authors have eliminated some self-citations. We want to tell the reviewer that the fact that there are so many self-citations is due to the fact that our research groups have been working for many years with biostimulants very similar to those in the manuscript to bioremediate soils contaminated by different organic xenobiotics. There are very few research groups working on this issue. For this reason, we consider the high number of self-citations to be normal. However, we hope that this is not an inconvenience for the work to be accepted for publication.

 

L14 manuscrpito? Is this a Spanish word? I think you mean manuscript! But, the word manuscript is not suitable to be used here, instead you can write (In this study, we …)

Corrected

 

L14 add , after manuscrpito

Corrected

 

L19 bacGram? What is this? I know it, but authors should define any abbreviations in the first mention to make the text clear and understandable for readers.

Revised

 

L20 what is Gram? Please define

Revised

 

L19-21 please add was before the word of decreased in this sentence. So, add it three times.

Corrected

 

L22-23 greater than what? Please add full meaning for text.

Corrected

 

L26 add sewage sludge as keyword

Word added

 

L33 remove these

Removed

 

L33-35 please cite these relevant ref: https://doi.org/10.1016/j.chemosphere.2012.07.031 

This reference has been included

 

L35 or in other places: please use sludge as a singular instead of using this as plural

Revised

 

L37-42 Please cite the relevant references for this text.

Revised

 

L43-46 Please cite this very relevant ref: https://doi.org/10.1016/j.resconrec.2019.104647

 

https://doi.org/10.1016/j.chemosphere.2006.12.019

References have been included in the text

 

L47-49 please add this relevant citation: https://doi.org/10.1016/j.biombioe.2013.07.021

This reference has been included

 

L55 what are these studies? Please cite two of the relevant or some of the above-cited references.

References are provided in the paragraph (Rodríguez-Morgado et al., 2015; Tejada et al., 2014).

 

L91 was NOT is

Corrected

 

L27 ha-1 make -1 as superscript

Corrected

 

L130-131 based on what you added 9.23 mg for each 300 g soil to get 2 kg per ha? Then, how many kg soils per ha do you think?

As we have commented at the beginning, we had an important mistake at the time of writing the manuscript. Regarding the kg of soil per hectare, the data requested is 2,010,000 kg of soil.

Keep in mind that the apparent density of the soil is 1.34 g/cm3 and that we took the first 15 cm of depth.

 

L133 BS1, BS2 and BS3??? What are these? Suddenly appeared, please define

Corrected

 

L157 why authors decided to analyze the diuron content in the soil at 2, 6, 10, 20, 35 and 55 days? Why was not done at 5,10,20,40,55 or …?

To answer this question, the authors want to highlight to the referee that we have worked with various biostimulants obtained mainly by enzymatic hydrolysis processes in the biochemical activity of the soil. The authors have seen that the most important changes occur in the first 10 days. This is because since these biostimulants are made up of low molecular weight proteins, they are quickly absorbed by soil microorganisms. Therefore, we think that in the early stages after applying biostimulants to contaminated soil, a faster response to the change in herbicide concentration in the soil can be observed. Consequently, this is the reason for determining the herbicide on the indicated days.

 

L163-170 revise the Statistical analysis, it was not well written.

Revised

 

Table 1: Why S, Ca and Pb are very high concentrations??

We have reviewed the data according to our laboratory notebook and they are correct

 

L201 please use p ≤0.05 NOT p <0.05 , correct this in other 9 places within your ms.

Corrected

 

Table 2, can be converted to Figure?

The authors have tried to convert all tables into figures. However, there is no optimal visualization. For this reason we have chosen to keep the tables. hope this is not a problem

 

Table 2: Why β-glucosidase activity was not reduced significantly as Dehydrogenase activity was reduced by increasing the period on incubation? The reason? This should be deeply explained in the discussion section.

The authors do not understand your question. All the enzyme activities analyzed decreased significantly in the soil contaminated with diuron with respect to the control treatment (without diuron). This decline occurs throughout the incubation process.

 

Discussion section has to be significantly improved, the authors have to make it deeper and explain the different mechanisms of their treatments in reducing or increasing their parameters instead of saying that our results were in line with this and this. If you say this, what is the new of your investigation?

We have rewritten the Discussion section according to your suggestions.

L343-344 common text, remove it. In conclusion, authors should focus on the most important finding at their work.

Deleted phrase

Reviewer 2 Report

Major revisions were required for the publication of this manuscript entitled “Use of biostimulants obtained from sewage sludge in the restoration of polluted soils by diuron. Effect on soil biochemical properties”.

The research conducted by the author is interesting but the presentation of the work is very poor. A lot of grammatical mistakes are there. The language throughout the manuscript is inadequate. Author's message is unclear.

1.     Improve the title of this manuscript.

2.     Rewrite the abstract of the manuscript. There are huge mistakes.

3.     Line 14: manuscripto?? Correct it.

4.     Line 19 bac Gram?? Please write it properly as gram positive or gram negative.

5.     Line 20 gram population?? Gram positive or negative. Please correct it.

6.     Line 21: The application of the three biostimulants to the soil decreased the concentration of diuron in the soil.?? On what basis authors concluded this. There is no information about the concentration and detection method of diuron no results explained. Only enzymatic results in abstract.

7.     Name of the family, bacteria and species should be in Italics in whole manuscript. Please correct

8.     Please include the NCBI accession number of bacterial isolates used in this study.

9.     Improve the introduction part with recent references from 2018-2022. Cover every section of this study in introduction part related to the microbial enzymes responsible for degradation process like laccase, dehydrogenase etc with mechanism. Authors may consider these works: https://doi.org/10.1007/s11756-021-00816-8, https://doi.org/10.3389/fmicb.2022.891870

10.  Please make a table for different treatments. Line 141-148.

11.  Where the soil treated with diuron under pot condition or field condition. Mention it clearly and some pictures related to this experiment.

12.  Please improve the heading of all tables.

13.  Explain the detailed methods for soil enzymes in material method section.

14.  Table 1: Organic matter standard error 17, 12, 11 and 19 check it. It is correct?? For Cu 11, 2.1, 15 and 10?? How it is varied in BS1?? Check it.

15.  Discussion section. Please include recent references related to your study.

Author Response

Major revisions were required for the publication of this manuscript entitled “Use of biostimulants obtained from sewage sludge in the restoration of polluted soils by diuron. Effect on soil biochemical properties”.

The research conducted by the author is interesting but the presentation of the work is very poor. A lot of grammatical mistakes are there. The language throughout the manuscript is inadequate. Author's message is unclear.

La investigación realizada por el autor es interesante pero la presentación del trabajo es muy pobre. The manuscript has now been reviewed by a native English speaker. we hope the grammatical problems have been fixed

 

 

Before beginning to answer the referees' questions, the authors want to comment on one matter.

Fortunately reviewer 1 made us a comment about the amount of soil used in the experiment and after reviewing the data we observed that we were confused with data from another experiment that we are carrying out with diuron.

In this experiment developed in this manuscript, the authors used 9.3 kg of soil for a dose of 2 kg/ha of diuron, with which 9.23 mg of the herbicide was added to this amount of soil.

We hope that this error of ours will not negatively influence the decision on the manuscript.

The authors want to make it clear that the data in the tables of the biological properties of the soil as well as those of the concentration of diuron in the soil are correct.

therefore we apologize for this great mistake that we have committed and that fortunately we have corrected

 

1. Improve the title of this manuscript.

The title of the manuscript has been revised

 

2. Rewrite the abstract of the manuscript. There are huge mistakes.

The manuscript has now been reviewed by a native English speaker

 

3. Line 14: manuscripto?? Correct it.

Corrected

 

4. Line 19 bac Gram?? Please write it properly as gram positive or gram negative.

Revised

 

5. Line 20 gram population?? Gram positive or negative. Please correct it.

Revised

 

6. Line 21: The application of the three biostimulants to the soil decreased the concentration of diuron in the soil.?? On what basis authors concluded this. There is no information about the concentration and detection method of diuron no results explained. Only enzymatic results in abstract.

Indeed, the application of the three biostimulants decreased the concentration of diuron in the soil. The abstract indicates that this decrease was greater when the biostimulant obtained by enzymatic hydrolysis was applied.

In the same way, it is true that there is no data that the reviewer suggests, but it is necessary to take into consideration that this section is the abstract one and that it has a word limit.

We think that the abstract gives a general idea of the manuscript. All the data requested by the reviewer are developed in the manuscript.

 

7. Name of the family, bacteria and species should be in Italics in whole manuscript. Please correct

Revised

 

8. Please include the NCBI accession number of bacterial isolates used in this study.

The authors do not understand your suggestion. We only use the Bacillus licheniformis ATCC21415 bacterium to carry out the fermentation process.

 

9. Improve the introduction part with recent references from 2018-2022. Cover every section of this study in introduction part related to the microbial enzymes responsible for degradation process like laccase, dehydrogenase etc with mechanism. Authors may consider these works: https://doi.org/10.1007/s11756-021-00816-8, https://doi.org/10.3389/fmicb.2022.891870

We have included in the Introduction section the mechanism of action of each enzymatic activity determined in the manuscript and the role of the study of soil microbial biomass

 

10. Please make a table for different treatments. Line 141-148.

Corrected

 

11. Where the soil treated with diuron under pot condition or field condition. Mention it clearly and some pictures related to this experiment.

The experiment was carried out in 15 kg pots. Sorry we do not provide photographs.

 

12. Please improve the heading of all tables.

Revised

 

13. Explain the detailed methods for soil enzymes in material method section.

Revised

 

14. Table 1: Organic matter standard error 17, 12, 11 and 19 check it. It is correct?? For Cu 11, 2.1, 15 and 10?? How it is varied in BS1?? Check it.

We have reviewed the data of the deviations corresponding to organic matter and although it is true that they are a bit high, they are correct. Regarding the Cu data, it is true that the value for BS1 is wrong. We put a comma in the figure when there really isn't a comma. We have also reviewed the deviations from said values for this parameter and they are correct according to our laboratory notebook.

 

15. Discussion section. Please include recent references related to your study.

We have rewritten the Discussion section. We have not been able to include more recent references. The reason for this is due to two main causes. First of all, there are very few (and not recent) studies about the effect of diuron on the biological properties of the soil. Secondly, we have not found manuscripts where biostimulants obtained by fermentation processes or enzymatic hydrolysis that affect the bioremediation of soils contaminated by diuron are used. Furthermore, we have not found any manuscripts that study the bioremediation of soils contaminated by herbicides using this series of biostimulants described in the manuscript that have been written by research groups other than ours.

 

 

Round 2

Reviewer 1 Report

Dear Authors

 

Thank you very much for revising your ms according to our comments. Now, the ms can be accepted.

 

Best Regards

Reviewer

Reviewer 2 Report

1 Authors have improved the manuscript.